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Mastitis in the dairy cow as revealed by field and laboratory tests Berry, John Coulter 1937

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MASTITIS I N THE DAIRY COW AS K2VEALED BY F I E L D AND LABORATORY TESTS . By-J o h n 0, B e r r y , B. S. A. , a n d F r a n k C. C l a r k , B. S. A. A T h e s i s S u b m i t t e d F a r The Degree O f MASTER OF SCIENCE I N AGRICULTURE I n The D e p a r t m e n t Of , ANIMAL HUSBANDRY, The U n i v e r s i t y o f B r i t i s h A p r i l , 1 9 3 7 . C o l u m b i a . ACKFOWLE DG-EMEWTB Ths authors wish to acknowledge the help and direction rendered by Professor H„ M. King, Head of the Department of Animal Husbandrys Dr» Blythe Eagles, Head of t h e Department of Dairying,, Miss 0. Okulitch-Assistant i n the Department of Dairying, and Dr, J", G. J e r v i s . Lecturer in Animal Pathol-ogy, in connection with the planning and de-velopment of thi s investigation, the provis-ion of equipment and f a c i l i t i e s for carrying out the various tests and for valuable guid-ance in the compilation of material. Thanks are due, also, to the dairy farm operators whose c ooperat ion has made this work possible. J . G . Berry. -B « Go GXsiZTIC«. TABLE! Off .OON.oai.TO-; INTRODUCTION: Scope and purpose of investigation. incidence and economic importance of mastitis. TYPES Off MASTITIS; EXPERIENCE .-Off THE AUTHORS WITH ffORMS OP  MASTITIS. Iip'EQTIQll " - " ATTITUDE Off DAIRYMEN IN GENERAL TO THE SUIT- ABILITY Off .MIL.H"EQR . ffLUID CONSUMPTION . VIEW Off INVESTIGATORS RE THE ABILITY Off MASTITIS INB'ECTED GO-WS TO RECOVER ff-RQM THE INffECTION;. EXPERIMENTAL; Procedure. Tests Employed to Detect Mastitis* Procedure ffoilowed i n Classifying Quarters witli Regard to Ma s t i t i s Infection. PRESENTATION Off RESULTS AND DISCUSSION: ' • • ' Explanations Relative to Presentation of Material. Explanations of Letters and Abbreviations Used i n Tables. Table No. 1 — Tabulated Results obtained from the Milk Samples of Cows Nos. 1 - 1 5 . Reaction to Dislac — Chlorine and Bacterial Content (with accompanying conclusion re Mastitis.Infection). Table No. 2 --Tabulated Results obtained from the Milk Samples of Cows Nos. 14 - 26. Reaction to Dislac -- Chlorine and Bacterial Content — Hotis Test, (with accompanying conclusion re Mastitis Infection). TABLE OF CONTENTS (continued) Table Ho. 3 — Tabulated Results obtained from the Milk Samples of Cows Nos. 2 7 - 5 3 . Reaction to Dislae -- Chlorine Con-tent -- Hotis Test (with accompany-ing conclusion re Mastitis Infection). Table ho. 4 "Case History" as related to "Conclusion re Mastitis Infection" Summary of Table ho. 4. Discussion of Table No. 4. Significance and Application of Material in Table ho. 4, CHLORINE CONTENT Off MILK SAMPLES AS RELATED  SO HOTIS TEST? ' ~ Table l o . j? — Chlorine Content of Milk "Samples as Related to Hotis Test. Discussion of Table l o . 5 . SUMMARY AID CONCLUSIONS: 1. MASTITIS IN THE DAIRY COW AS REVSALSP BY • PTMLPA AND1ABQR,\T OR Y TESTS . INTRODUCTION. M a s t i t i s ( c o n t a g i o u s g a r g e t ) I s a d i s e a s e o f t h e u d d e r o f d a i r y c a t t l e w h i c h has be e n known t o e x i s t t o a c o n -s i d e r a b l e e x t e n t i n t h e d a i r y h e r d s o f B r i t i s h Columbia,, e v e r s i n c e t h e v e r y e a r l y days o f t h e d a i r y i n g i n d u s t r y . P r e v i o u s t o t h e i n v e s t i g a t i o n h e r e i n p r e s e n t e d ; no a t t e m p t h a d be e n made t o a s c e r t a i n t h e i n c i d e n c e and n a t u r e o f t h i s i n c r e a s i n g l y i m p o r t a n t d i s e a s e I n t h e areas, o f t h i s P r o v i n c e where d a i r y i n g c o n s t i t u t e s a m a j o r i n d u s t r y . I n t h e P a . l l o f 193.6s t h e a u t h o r s commenced an i n v e s -t i g a t i o n i n t o m a s t i t i s c o n d i t i o n s i n s e v e r a l r e a . d i l y a v a i l a b l e d a i r y h e r d s o f t h e L o w e r P r a s e r V a l l e y , I n t h i s a r e a , s i t -u a t e d i n t h e ex t r e m e S .\7. p a r t o f t h e P r o v i n c e , t h e d a i r y i n g I n d u s t r y i s v e r y well established. A f e w h e r d s i n t h e n e i g h -b o r h o o d o f K a m l o o p s a . C i t y i n t h e I n t e r i o r s , were a l s o s e l e c -ted, f o r s t u d y * The p r i m a r y o b j e c t o f t h i s i n v e s t i g a t i o n h a s be e n t o d e t e r m i n e t h e u d d e r c o n d i t i o n and t h e n a t u r e o f t h e m i l k s e c r e t e d b y cows w h i c h have s u s t a i n e d t e a t o r u d d e r i n j u r i e s o r a known i n f e c t i o n f r o m m a s t i t i s a t some t ime. and s u b s e -q u e n t l y r e c o v e r e d t h e a b i l i t y t o s e c r e t e a p p a r e n t l y n o r m a l * m i l k . The m a j o r i t y o f t h e cows s e l e c t e d f o r t e s t i n g , t h e r e -* A p p a r e n t l y n o r m a l -~ r e f e r r i n g t o m i l k t h a t a p p e a r s n o r m a l f r o m a p h y s i c a l e x a m i n a t i o n , o r , r e f e r r i n g t o cows t h a t h a v e had no known t e a t o r u d d e r i n j u r i e s , a n d no known p r o d u c t i o n o f o t h e r t h a n a p p a r e n t l y n o r m a l m i l k e 2-. f o r e . had s u c h c a s e h i s t o r i e s * , and were a t t h e t i m e o f samp-l i n g 5 s e c r e t i n g m i l k t h a t , on p h y s i c a l e x a m i n a . t i o n s a p p e a r e d n o r m a l . The a u t h o r s w i s h t o p o i n t o u t t h e n e c e s s i t y o f k e e p -i n g t h i s b a s i s o f s e l e c t i o n o f cows f o r t e s t i n g i n mind when a s t u d y i s made o f t h e d a t a h e r e i n . A t t h e same t i m e , b e c a u s e t h e h e r d s f r o m w h i c h cows were s e l e c t e d w e re u n d e r b e t t e r - t h a n -a v e r a g e management:«, a n d b e c a u s e b o t h a p p a r e n t l y n o r m a l cows and and cows s e c r e t i n g o b v i o u s l y a b n o r m a l m i l k w e re l a r g e l y a v o i d e d i t i s f e l t t h a t t h i s i n v e s t i g a t i o n i s o f v a l u e i n i n d i c a t i n g t h e i n c i d e n c e o f m a s t i t i s i n B r i t i s h C o l u m b i a ' s d a i r y h e r d s * M a s t i t i s has l o n g been r e c o g n i z e d a s one o f t h e d i s e a s e s c a u s i n g s e v e r e economic l o s s t o d a i r y f a r m e r s . The f o l l o w i n g f e w q u o t a t i o n s ( l ) w i l l s e r v e t o g i v e a g e n e r a l i d e a o f t h e i n c i d e n c e and s i g n i f i c a n c e o f t h e d i s e a s e t D r . t i d a l 1, M e d i c i n e O b s t e t r i c o f t h e C o r n e l l C o l l e g e o f V e t e r i n a r y M e d i c i n e , 1931s s t a t e s : "ho one c a n e s t i m a t e t h e e x a c t e c o n o m i c a l l o s s t o t h e D a i r y I n d u s t r y - c a u s e d by m a s t i t i s , , I n a 11 p r o b a b i l i t y i t w o u l d e x c e e d t h a t b r o u g h t a b o u t by t u b e r -c u l o s i s o r by a b o r t i o n . The o f f i c e r s o f t h e H e a l t h D e p a r t m e n t s and a l s o h e r d - o w n e r s a r e more a n d more a s t o n i s h e d by t h e r e l a -t i v e l y sma11 i m p o r t a n c e t h a t has been a t t a c h e d t o t h i s d i s e a s e , " * Case h i s t o r y — r e l a t i n g t o t h e c o n d i t i o n s p r e v i o u s t o t h e t i m e o f d r a w i n g m i l k s a m p l e s , o f t h e t e a t s , u d d e r , a n d m i l k o f a cow. 3 *• Ostertag, President of the Veterinary Society of Germany, i n his report to the International Congress of Vet-erinary Medicine held, i n London, 1 9 3 1 > said that next to -tuberculosis: "Streptococcic Mastitis is the most wide-spread and important disease to dairy herdsmen." The North American Veterinarian 1 0 = I929, p»59:~ "Mastitis i s an ever-increasing menace to the Milk Industry. The development of this disease throughout the country appears to be increasing more rapidly than the development of the milk industry i t s e l f . There is not one animal disease to-day which i s drawing so much deserved attention from vet-erinarians ." Seelemann i n a publication of the Milk Research Institute of Prussia, in K i e l , 1 9 3 2 , gives the following figures: 170 herds t o t a l l i n g 769I heads of cattle were exam-ined and 2539 were found to be infected with mastitis, v i z . 33%? 14-59 were suspected ( 1 9 ^ ) , leaving only 4-8% as healthy. At the Annual Veterinary Convention held in 1 9 3 1 at the College of Veterinary Medicine at Cornells Holfort, Chief Veterinarian of Borden's farm, found that for the State of New York, out of 1 , 3 3 0 , 0 0 0 cows, 57s190 are annually elim-inated from production through mastitis. This figure repre-sents a loss of 3 » 2 3 6 m i l l i o n pounds of milk, or roughly 49 m i l l i o n dollars per annum, the total loss being 72' m i l l i o n dollars for one year. 4,. T. G. Minett, Director of the Research Institute of Animal Pathology of the Royal Veterinary College of London, stated in 1 9 3 0 : "850 cows were examined and 39% were found to be infected with streptococcus mastitidis." Case (2) a practising veterinarian,in an address presented at the 7 3 r d A n n u a l Meeting of the American Veterin-ary Medical Association, Columbus, Ohio, i n August 1936, stated: "At the present time, mastitis causes more l o s s to dairymen than a l l of the other diseases of dairy cattle com-bined. There are many factors which contribute to this loss, one of the chief ones being the decreased value of the affec-ted cows. This i s the direct result of their producing milk deficient i n both quantity and quality." L. A. Rogers ( 3 ) s Chief, Div i s i o n of Dairy Research Laboratories, Bureau of Dairy Industry, U. S. Department of Agriculture, Washington, D.C, i n an address at the Ninth Annual Institute of Dairying at the State College of Washing-ton, Pullman, Washington, March 1 9 3 ° , stated: "It i s estim-ated that f u l l y one-third of the cows producing milk for the City trade are more or less affected by mastitis and i n some cases the incidence may be as high as two-thirds. It i s gen-e r a l l y recognised as a more serious disease from the economic standpoint than tuberculosis or infectious abortion." Quoting again from Rosell -- "In studies I have made in Canada during the l a s t f i v e years under the auspices of the National Research Council and the Department of Agri-\culture of Quebec, I have found that from 1838 cows tested i n the separate milk of each quarter, 34.6% were a f f l i c t e d with mastitis. In testings I have made in the United States, s'orne of them in the laboratories of the Bureau of Animal Indus-try, i n collaboration with Dr. W. T. M i l l e r , the percentage of cows suffering from this disease was found to be s t i l l larger -- The number of individual cows we have found infected with mastitis in the separate herds i n Canada., was between 19% (the lowest percentage) to 97% of the cows tested." It i s readily evident from the above opinions (which are representative of the many that have been expressed) that mastitis i s a very important disease, p a r t i c u l a r l y from the point of view of those d i r e c t l y connected with the dairying industry. In view of the huge t o l l i t takes from our national wealth and i n view of the public health aspect of the unques-tioned u t i l i z a t i o n of milk from mastitis infected- cows for human consumption, the problem i s of universal concern. TYPES OF MASTITIS Various types of mastitis i n f e c t i o n have been men-tioned by a number of investigators. There appears to be some lack of uniformity i n terminology. Emphasis has been placed on mastitis caused by streptoccus mastitidis organisms. In-fection of this nature has been c l a s s i f i e d into three types by Hucker (4). He mentions c l i n i c a l mastitis, stating "This type of mastitis i s generally referred to as the acute, active type and is r e l a t i v e l y uncommon. The cow i s generally pros-6. trated, there is an intense l o c a l inflammation, l i t t l e or no milk can he secured from the affected quarter, toxemia i s evident, temperature i s high, and the i n f e c t i o n generally ends f a t a l l y or with a, loss of the quarter. It i s estimated that about eight cows per thousand per year have this acute type of mastitis, "The organisms responsible for this type of masti-t i s are usually of a toxin-producing type. Generally, toxin-producing streptococci are associated with c l i n i c a l mastitis, although i t may he caused hy colon organisms and certain types of virulent micrococci (staphylococci). One interesting observation i n connection with the c l i n i c a l type of mastitis i s the f a c t that i t rarely spreads through the herd. It occurs in isolated instances and, although the cow has been i n the milking l i n e previous to infection, adjoining cows rarely become infected. It i s also of interest to note that at the height of the i n f e c t i o n the organisms are r e l a t i v e l y d i f f i c u l t to isolate from the secretions from the affected quarter." The second type mentioned i s s u b - c l i n i c a l (sub-acute, chronic, latent, etc.) mastitis. "This i s the most prevalent type of mastitis i n New York State dairy herds. The ind.ivid.ual may have no external appearance of an infec-tion, the milk may remain normal i n appearance, and the cow give no evidence of abnormal conditions. Even l o c a l inflam-mation may be absent and she may remain i n the milking l i n e as a perfectly normal animal. On the other hand, strepto- • 7. cocci may "be found in the milk, the leucocyte count may be high, the quarter may show hardened areas or scar tissue, flakes may appear on the s t r i p cup, and an alkaline reaction may.be obtained with the brom thymol blue test. The incidence of this type of mastitis i n hew York State i s r e l a t i v e l y high. A survey of a, large number of herds indicates that from 20 to 90 per cent of the individuals i n the. herd may be infected. The organism causing the infection i s probably a streptococ-cus. Its virulence i s extremely low. The organisms probably gain entrance through the teat canal and a chronic infect i o n takes place. The in f e c t i o n progresses and the quarter becomes gradually f i l l e d with scar tissue which may be eas i l y demon-strated, by manual manipulation. This type of mastitis i s of great economic significance in Hew York State. "Both the c l i n i c a l and the chronic types may occas-i o n a l l y spread rapidly through the herd and become infectious or contagious, or they may occur as isola t e d sporadic cases." The t h i r d type mentioned by Hucker i s septic sore throat mastitis: "This type of mastitis should be d i s t i n -guished sharply from ordinary bovine mastitis, due to the fact that i t is caused by a different type of organism and has an ent i r e l y different sanitary significance. Septic sore throat mastitis is thought to be caused hy a human pathogenic strep-tococcus. This type of streptococcus has i t s origin i n the throats of the milkers or milk handlers and o r d i n a r i l y gains admission to the udder through an abrasion of the skin or teat 8 , canal. The type of mastitis which i s set up is not severe i n nature, shows only a small amount of l o c a l inflammation, with l i t t l e or no systemic reaction in the cow, and the milk may "b of normal appearance. The use of this milk, i f not pasteur-ized, generally results i n a case of human septic sore throat Rosell ( l ) , i n discussing the indolence of various types of mastitis, states that the ordinary chronic or sub-c l i n i c a l forms of mastitis constitute 9 5 % or more of the mas-t i t i s a f f l i c t i n g the milk industry today, and that only streptococci (Str. mastitidis or agalactiae) are secured i n e t i o l o g i c a l connection with the disease. Rogers (3) stated i n the Spring of 1 9 3 6 , that the causal organism of mastitis has not been p o s i t i v e l y e s t a b l i -shed, and that apparently i t may be caused by staphylococci, by colon bacteria, and by several members of the streptococ-cus group. He said further that nearly a l l investigators would agree that 8 0 or 9 0 % of the cases of chronic mastitis are caused by a single organism, a streptococcus known some-times as Str. mastitidis, but more frequently as Str. agala.c-tiae . A somewhat different opinion i s expressed by Case (2) who has found i n his practice that staphylococci cause a low-grade infection i n 39% of the cases of mastitis. 9« EXPERIENCE OE THE AUTHORS WITTS. FORMS OE MASTITIS INFECTION • The authors are acquainted with a. herd which had never known a case of mastitis u n t i l following the purchase of a dry cow at a consignment sale,. After freshening this cow produced clotty miIk and the mastitis infe c t i o n thus intro-duced, was widely spread after a year or two throughout this previously healthy herd. In another herd (with which one of the authors has been closely associated for some years) both the acute and the chronic forms of ma.stitis have been present more or less con-tinuously. Some twelve years, ago a very severe outbreak of the disease occurred in this herd. Previous to that time the cows in the herd had been r e l a t i v e l y free of the disease, but following the purchase of a number of animals the acute form of mastitis developed. One of the f i r s t few cows to become infected developed a high temperature, a shivering condition, her udder became highly inflamed, and she died, within forty-eight hours. Death did not result In the other cases at or since that time. A number of the o r i g i n a l l y infected animals, were, pure-bred cows of considerable value for reproductive pur-poses, and hence were treated for the trouble and retained in the herd „ Some of them recovered the ab i l i t j r to secrete ap-parently normal milk from the infected quarters although frequently in lessened quantity. These cows were also more subject than the healthy animals of the herd to the occasional 10 . s e c r e t i o n , f r o m t i m e t o t i m e , o f s l i g h t l y c l o t t y m i l k , A few GOWS temporarxly s and o t h e r s permanently, l o s t t h e use o f one or.more q u a r t e r s . We have f o u n d i n o u r e x p e r i e n c e t h a t q u i t e a p p r e -c i a b l e numbers o f d a i r y cows may ha v e a c u t e a t t a c k s o f m a s t i -t i s ™- t h e m i l k becomes c l o t t y , s e r o u s , and f r e q u e n t l y h a s a d i s a g r e e a b l e o d o r . The a f f e c t e d q u a r t e r s q u i c k l y d r y up and r e m a i n b l i n d * d u r i n g t h e b a l a n c e o f t h e l a c t a t i o n * A t t h e s t a r t o f t h e n e x t l a c t a t i o n s u c h q u a r t e r s n o t i n f r e q u e n t l y p r o d u c e m i l k t h a t a p p e a r s n o r m a l f r o m a p h y s i c a l examination* I t i s o u r e x p e r i e n c e , h o w e v e r , t h a t q u a r t e r s once i n f e c t e d w i t h an a c u t e o r a, m i l d a t t a c k o f t h e d i s e a s e ( a s i n d i c a t e d by t h e s e c r e t i o n o f c l o t t y o r s e r o u s m i l k ) a r e u s u a l l y s u b j e c t t o r e c u r r e n t a t t a c k s f r o m t i m e t o t i m e . T h i s i s e s p e c i a l l y t r u e i f t h e animals a r e s u b j e c t e d t o h i g h p r o t e i n f e e d i n g s s u d den c h a n g e s i n t h e i r r a t i o n s , o r some p h y s i c a l d i s t u r b a n c e s u c h a s e x p o s u r e t o e x c e s s i v e c o l d . I n some c a s e s t h e f i r s t i n d i c a t i o n o f t r o u b l e i n a; cow's u d d e r h a s been a t e a t i n j u r y o r a s l i g h t l y i n f l a m e d c o n d i t i o n i n a q u a r t e r due p o s s i b l y t o a c h i l l o r some o t h e r p h y s i c a l cause.. I n o t h e r c a s e s a s l i g h t l u m p i n e s s ( f i b r o s i s ) i s d e t e c t e d i n a p r e v i o u s l y n o r m a l q u a r t e r , , These c o n d i t i o n s o f t e n d e v e l o p g r s . d u a l l y and d u r i n g t h e e a r l y s t a g e s t h e m i l k p r o d u c e d i s a p p a r e n t l y n o r m a l . E v e n t u a l l y , h o w e v e r . a f l a k e y ^ R e f e r r i n g t o a q u a r t e r t h a t has l o s t t h e f u n c t i o n o f m i l k s e c r e t i o n * 11. c l o t t y ^ o r s e r o u s c o n d i t i o n may be d e t e c t e d i n t h e m i l k a s drawn* The p r o d u c t i o n o f a b n o r m a l m i l k b y a q u a r t e r t h a t h a s been s l i g h t l y i n f e c t e d f o r some t i m e i s u n d o u b t e d l y h a s t e n e d by u n f a v o r a b l e e n v i r o n m e n t a l c o n d i t i o n s o r p h y s i c a l c a u s e s . On t h e o t h e r hand, a s w i l l be shown by t h e work p r e s e n t e d i n t h i s i n v e s t i g a t i o n , m a s t i t i s i s f o u n d i n i n s t a n c e s i n w h i c h no i n d i c a t i o n o f t h e d i s e a s e has e v e r been o b s e r v e d , e i t h e r i n t h e m i l k o r i n t h e p h y s i c a l c o n d i t i o n o f t h e u d d e r . ATTITUDE 03? DAIRYMEN I N GENERAL TO THE S U I T A B I L I T Y OE MILK FOR FLUID CONSUMPTION We h a v e been d e a l i n g t h u s f a r w i t h v a r i o u s t y p e s and d e g r e e s o f u d d e r i n f e c t i o n , l a r g e l y f r o m t h e p o i n t o f v i e w o f t h e i n v e s t i g a t o r , We now w i s h t o p r e s e n t t h a t w h i c h . i n o u r e x p e r i e n c e a n d o p i n i o n , i s t h e a t t i t u d e o f t h e a v e r a g e d a i r y m a n . M i l k i s , o f c o u r s e , d i s c a r d e d a s u n f i t f o r human c o n s u m p t i o n when i t s p h y s i c a l a p p e a r a n c e i s a b n o r m a l t h a t i s , , when i t i s c l o t t y , w a t e r y , o f f - c o l o r , o r when i t has a d i s a g r e e a b l e o d o r * I n some c a s e s ( b u t i n o u r o p i n i o n t h e s e a r e t h e e x c e p t i o n s ) m i l k f r o m q u a r t e r s t h a t a r e s l i g h t l y s w o l l e n o r lumpy ( i n d i c a t i n g t h e p r e s e n c e o f s c a r t i s s u e ) a l s o f a i l s t o f i n d i t s way I n t o t h e n o r m a l c h a n n e l s o f u t i l i z a t i o n . However, s h o r t l y a f t e r cows w h i c h h a v e had some i n f e c t i o n , r e c o v e r t h e a b i l i t y t o s e c r e t e m i l k t h a t i s a p p a r e n t l y norma.!, s u c h m i l k i s marketed, f o r f l u i d c o n s u m p t i o n . F u r t h e r , s u c h cows a r e r e g a r d e d a s h a v i n g r e c o v e r e d f r o m m a s t i t i s i n f e c t i o n 1 ? ( a t l e a s t t e m p o r a r i l y I f n o t p e r m a n e n t l y ) and a r e t r e a t e d i n t h e h e r d a s n o r m a l cows. We h o l d t h a t s u c h i s t h e v i e w and t h e p r o c e d u r e o f t h e g r e a t m a j o r i t y o f d a i r y farmers» VTE¥ 03? INVESTIGATORS RE THE A B I L I T Y OE MASTITIS INFECTED COWS TO RECOVER EROM THE INFECTION* R o g e r s ( 3 ) s t a t e d i n M a r c h , 1 9 3 6 , "Our o b s e r v a t i o n s i n d i c a t e t h a t w h i l e a cow may n e v e r r e c o v e r f r o m a n i n f e c t i o n i n t h e s e n s e t h a t t h e a f f e c t e d t i s s u e d o e s n o t r e t u r n t o n o r m a l , t h e change i n t h e m i l k I s n o t p e r m a n e n t and when t h e i n f l a m m a t i o n s u b s i d e s i t r e t u r n s t o i t s n o r m a l c o n d i t i o n " f t D r . G. J . H u c k e r o f t h e New Y o r k S t a t e A g r i c u l t u r a l E x p e r i m e n t S t a t i o n , i n a l e t t e r t o t h e a u t h o r s i n M a r c h , 1 9 3 7 , e x p r e s s e d t h e o p i n i o n t h a t an u d d e r "once i n f e c t e d a l w a y s i n -f e c t e d " . F r o m a s t u d y of t h e l i t e r a t u r e r e l a t i n g t o m a s t i t i s i n d a i r y c a t t l e t h e a u t h o r s have f o u n d l i t t l e r e f e r e n c e t o t h i s p h a s e o f t h e p r o b l e m . M o s t o f t h e w o r k t h a t has been done a p p e a r s t o r e l a t e t o a s t u d y o f t h e more a d v a n c e d s t a g e s o f t h e d i s e a s e . As w i l l be s e e n f r o m a p e r u s a l o f t h e e x p e r -i m e n t a l p r o c e d u r e i n t h i s i n v e s t i g a t i o n t h e a u t h o r s a r e a t t e m p t i n g t o t h r o w some a d d i t i o n a l l i g h t on t h e m i l d e r c a s e s o f m a s t i t i s i n f e c t i o n ( t h a t i s where t h e p h y s i c a l n a t u r e o f t h e m i l k a p p e a r s n o r m a l . ) . EXPERIMENTAL The cows s e l e c t e d f o r e x p e r i m e n t a l p u r p o s e s may be g r o u p e d I n t o t h e f o l l o w i n g c l a s s e s ? 1. A p p a r e n t l y n o r m a l cows (no i n j u r i e s and no p h y s i c a l l y a b n o r m a l m i l k p r o d u c e d a t any t i m e , i n c l u d i n g t i m e o f s a m p 1 i n g ) . 2* Cows w i t h i n j u r i e s t o u d d e r o r t e a t s b u t w h i c h have n o t been known t o p r o d u c e o t h e r t h a n a p p a r e n t l y n o r m a l m i l k . 3. Cows w i t h a m a s t i t i s h i s t o r y ( h a v i n g a t some t i m e p r o -duced p h y s i c a l l y a b n o r m a l m i l k ) , b u t w h i c h a t t i m e o f iSampling were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k f r o m a l l f o u r q u a r t e r s * 4, Cows w i t h a m a s t i t i s h i s t o r y w h i c h a t t i m e o f s a m p l i n g were s e c r e t i n g m i l k t h a t a p p e a r e d p h y s i c a l l y a b n o r m a l * P r o c e d u r e ; M i l k s a m p l e s were drawn f r o m i n d i v i d u a l q u a . r t e r s i n t o s t e r i l e g l a s s c o n t a i n e r s , and t a k e n t o t h e l a b o r a t o r y f o r t e s t i n g . P r i o r t o t a k i n g t h e s a m p l e s f l a n k s and u d d e r s o f t h e cows were c l e a n e d , washed, and d i s i n f e c t e d . The f i r s t m i l k was d i s c a r d e d and t h e P i s l a c T e s t t h e n a p p l i e d * The m i l k s a m p l e s were t h e n p r o c u r e d . T h i s f i e l d w o r k was done a t e i t h e r t h e r e g u l a r noon o r e v e n i n g m i I k i n g s , and t h e l a b o r a -t o r y t e s t i n g was a c c o m p l i s h e d t h e f o l l o w i n g forenoon„ The p h y s i c a l c o n d i t i o n of t h e u d d e r and o f t h e m i l k was n o t e d a t t h e t i m e o f d r a w i n g t h e s a m p l e s . The c a s e h i s t o r y o f i n d i v i -d u a l cows r e l a t i n g t o t h e number o f months f r e s h a t t h e t i m e o f s a m p l i n g , , t o u d d e r o r t e a t i n j u r i e s , and t o t h e s e c r e t i o n o f a b n o r m a l m i l k ( i f a n y ) was o b t a i n e d a t t h i s t i m e . 14 » T e s t s E m p l o y e d t o D e t e c t M a s t i t i s ; P h y s i c a l E x a m i n a t i o n o f t h e U d d e r - - B y m a n i p u l a t i n g t h e q u a r t e r s o f t h e u d d e r t h e p r e s e n c e o r a b s e n c e o f n o d u l e s o-r l u m p l n e s s was d e t e r m i n e d . The o c c u r r e n c e o f s u c h i n d i c a t e s a f o r m a . t i o n o f s c a r o r c o n n e c t i n g t i s s u e ( f i b r o s i s ) i n p l a c e o f t h e n o r m a l s e c r e t i n g t i s s u e . P h y s i c a l E x a m i n a t i o n o f t h e M i l k The c o n d i t i o n o f t h e m i l k was no t e d a t t h e t ime o f d r a w i n g t h e s a m p l e s . A s l i g h t f l a k e y n e s s w h i c h m i g h t be p r e s e n t a n d p a s s e d u n n o t i c e d was i n v a r i a b l y d e t e c t e d by t h e P i s l a c T e s t . The D i s l a c T e s t •— The D i s l a c T e s t h a s t h e a d v a n t a g e o f c o m b i n i n g two t e s t s i n one. ( a ) t h e s t r i p - c u p t e s t u s e d i n d e t e c t i n g c l o t s i n mi l k , , (b) t h e t e s t w i t h b r o m - c r e s o l p u r p l e i n d i c a t o r u s e d f o r d e t e r m i n i n g t h e d e g r e e o f a l k a l i n i t y o f t h e m i l k b y t h e p r o d u c t i o n o f sha.des o f b l u e , v a r y i n g f r o m s l a t e g r e y ( n o r m a l ) t o a deep p u r p l e ( e x t r e m e a b n o r m a l ) , T h i s t e s t was c a r r i e d o u t by a l l o w i n g a j e t o f m i l k , d i r e c t f r o m t h e u d d e r , t o p a s s t h r o u g h a m u s l i n d i s c i m p r e g -n a t e d w i t h b r o m - c r e s o l p u r p l e i n d i c a t o r . The m u s l i n r e t a i n s a n y c l o t s p r e s e n t i n t h e m i l k , and i t c h a n g e s i n c o l o r f r o m y e l l o w t o p u r p l e i f t h e m i l k I s a l k a l i n e . The c h a n g e s i n c o l o r were n o t e d i m m e d i a t e l y b e c a u s e t h e c o l o r g r a d u a l l y d a r k e n s ( e v e n i n a w e l l v e n t i l a . t e d s t a b l e ) due t o t h e p r e s e n c e i n t h e a i r o f t r a c e s o f ammonia, A sla.te-g.rey c o l o r i s c o n s i d e r e d a n o r m a l r e a c t i o n . A d e e p e r c o l o r tha.n t h i s i s i n d i c a t i v e t h a t t h e m i l k may be a b n o r m a l e x c e p t i n t h e c a s e o f cows w h i c h a r e 1?. well advanced i n their lactation periods. Such cows gener-a l l y give a, more alkaline milk and hence a deeper color does not necessa r i l y indicate the presence of mastitis i n f e c t i o n . The appearance of clots on the muslin disc i s , of course, a definite indication of the presence of mastitis. Almost i n -variably , when clots appear the color reaction i s a dark purple« Not infrequently, however, i n our experience quite definite color reactions are obtained without the appearance of flakes or c l o t s , Plate Count Determination, of Ba c t e r i a l Content — A 1 j100 d i l u t i o n of the milk sample from each quarter was made, and 1 cc, of the d i l u t i o n plated on purple lactose agar* The plates were incubated at 3.2° 0• and. the number of colonies present was determined a f t e r twenty-four and forty-eight hours* In some instances circumstances made i t necessary to take readings aft e r other periods of incubation* In a l l cases, however, the time period is indicated in the tables of results. Plating of milk samples to determine bacterial con-tent was proceeded with u n t i l February 8th, a f t e r which date this test was discontinued. ¥ith the f i r s t use of the Hotis Test (see below) i t was f e l t that the plate count method, us-ing purple lactose agar, was. not of pa r t i c u l a r use i n the present work,.. Colonies were readily discernible in the Hotis tubes i f Streptococcus Mastitidis was present, and the presence or absence of this organism was a l l that was required regard-ing the bacteria present in a milk sample. l 6 h A l s o , due t o r e c e n t r e s u l t s (5) p u b l i s h e d on the d e t e r m i n a t i o n o f t h e b a c t e r i a l c o n t e n t o f m i l k b y p l a t i n g , i t was f e l t t h a t t h e p l a t e c o u n t method was r a t h e r u n r e l i a b l e due t o t h e d i f f i c u l t y o f e s t a b l i s h i n g a u n i f o r m l y a c c u r a t e i n t e r p r e t a t i o n * C h l o r i n e C o n t e n t B e t e r m i n a t i o n — The f o l l o w i n g method f o r t h e d e t e r m i n a t i o n o f c h l o r i n e i s t h a t u s e d by Ho s e l l (l) t "40 c c . o f d i s t i l l e d w a t e r a d d e d t o 10 c c . o f m i l k I n a s m a l l beaker;. 8 t o 10 d r o p s o f a 10% s o l u t i o n o f p o t a s s i u m chrornate a r e a d d e d t o g i v e a c a n a r y y e l l o w c o l o r t o t h e d i l u t e d m i l k , h/10 s i l v e r n i t r a t e s o l u t i o n i s r u n i n f r o m a b u r e t t e u n t i l t h e f i r s t p e r m a n e n t deep o r a n g e c o l o r a p p e a r s " . I n t h e a u t h o r s ' e x p e r i e n c e t h e f i r s t p e rmanent c o l o r change was f o u n d t o be f r o m a p a l e y e l l o w t o a p a l e o r a n g e . A c c o r d i n g l y , . h/10 s i l v e r n i t r a t e was t i t r a t e d i n t o e a c h m i l k sample u n t i l t h e l a t t e r d e g r e e o f c o l o r a t i o n was r e a c h e d , The number o f c c . o f s i l v e r n i t r a t e n e c e s s a r y t o p r o -duce, t h i s change m u l t i p l i e d by t h e m o l e c u l o r w e i g h t o f c h l o r i n e («03546) g i v e s , t h e c h l o r i n e c o n t e n t i n grams p r e s e n t i n 100 c c , mi I k , R o s e l l ( l ) s t a t e s t h a t " I f more t h a n 4 c c , o f h/10 s i l v e r n i t r a t e a r e r e q u i r e d t o n e u t r a l i z e t h e c h l o r i n e i n 10 c c . m i l k , m a s t i t i s i s p r e s e n t . " The same a u t h o r ( l ) m e n t i o n s t h a t "a c o n t e n t o f more t h a n Q.14% o f c h l o r i n e i s a l w a y s r e -l a t e d t o p a t h o l o g i c a l c h a n g e s i n t h e u d d e r , " and t h a t " a m i l k h a v i n g more t h a n 0.13% o r 0.14%' o f c h l o r i n e s h o u l d be cons i d -e r e d a b n o r m a l " . A t t h e t e n t h A n n u a l I n s t i t u t e o f D a i r y i n g h e l d a t t h e S t a t e C o l l e g e o f W a s h i n g t o n i n t h e S p r i n g o f 1937s D r . H e i s e r o f O h i o S t a t e U n i v e r s i t y , , w h i l e d i s c u s s i n g v a r i o u s t e s t s f o r t h e d e t e c t i o n o f m a s t i t i s , spoke f a v o r a b l y o f t h e c h l o r i n e t e s t a n d i n d i c a t e d tha,t a c h l o r i n e c o n t e n t g r e a t e r t h a n .15% was i n d i c a t i v e o f m a s t i t i s i n f e c t i o n . I n r e f e r e n c e t o t h e i n t e r p r e t a t i o n o f t h e c h l o r i n e t e s t s H u c k e r ( 4 ) makes t h e f o l l o w i n g s t a t e m e n t — "Under g e n -e r a l c - o n d i t i o n s i t i s assumed t h a t a l l m i l k s h o w i n g more t h a n 0.14 p e r c e n t c h l o r i d e s has b een d e r i v e d f r o m i n f e c t e d u d d e r s . I n t h e a b b r e v i a t e d t e s t t h e r e a g e n t s a r e so a d j u s t e d t h a t a y e l l o w c o l o r d e v e l o p s i n t h e m i x t u r e i f t h e c o n c e n t r a t i o n o f c h l o r i d e s i s g r e a t e r t h a n t h i s amount, "The p r o p e r i n t e r p r e t a t i o n o f t h e r e s u l t s o f t h i s t e s t i s somewhat d i f f i c u l t . I n v e s t i g a t o r s d i s a g r e e a s t o t h e n o r m a l c h l o r i d e c o n t e n t o f m i l k , From g e n e r a l e x p e r i e n c e , h o wever, i t h a s been assumed t h a t t h e n o r m a l c h l o r i d e c o n t e n t o f m i l k - v a r i e s f r o m 0.09 t o 0.14 p e r c e n t . C e r t a i n o b s e r v e r s have e l e c t e d t o c o n s i d e r a l l m i l k w i t h a. c h l o r i d e c o n t e n t g r e a t e r t h a n 0.3-5 p e r c e n t a s c o m i n g f r o m i n f e c t e d u d d e r s " * I n t h e p r e s e n t w o r k , i f 4,3 c c . o r more o f IT/10 s i l v e r n i t r a t e were r e q u i r e d t o n e u t r a l i z e t h e c h l o r i n e i n 10 c c , m i l k , t h e a u t h o r s c o n s i d e r e d t h a t m a s t i t i s was i n d i c a t e d The use o f 4.3 c c . o f h/10 s i l v e r n i t r a t e i n d i c a t e s a c h l o r i n e c o n t e n t o f .1524 grams i n 100 c c . m i l k . 18. The_ H o t i s T e s t f o r j h e D e t e c t i o n o f M a s t i t i s S t r e p t o c o c c i i n M i l k . — The H o t i s T e s t f o r d e t e c t i n g m a s t i t i s s t r e p t o c o c c i I n m i l k was d e v e l o p e d by R. P. H o t i s o f t h e U n i t e d S t a t e s B u r e a u o f P a i r y I n d u s t r y , W a s h i n g t o n , D, C», d u r i n g 193.3 " 1 9 3 ? 5 and i s c a l l e d t h e H o t i s T e s t i n r e c o g n i t i o n o f h i s w o r k , Mr. H o t i s d i e d i n J u l y , 1935? a n - ^* ^» M i l l e r o f t h e P a t h o l o g i c a l D i v i s i o n , B u r e a u o f A n i m a l I n d u s t r y , p r e p a r e d t h e m a t e r i a l f o r p u b l i c a t i o n . I n A u g u s t , 1936", C i r c u l a r No. 400, "A S i m p l e T e s t f o r D e t e c t i n g M a s t i t i s S t r e p t o c o c c i i n M i l k " ( 6 ) was ma.de a v a i l a b l e f o r d i s t r i b u t i o n by t h e U n i t e d S t a t e s D e p a r t m e n t o f A g r i c u l t u r e . I n v i e w o f t h e fa.ct t h a t t h i s i s a r e l a t i v e l y new m a s t i t i s t e s t , a nd b e c a u s e t h e a u t h o r s have compared, and c o r r e l a t e d i t i n a number o f ways w i t h t h e more e s t a b l i s h e d . " C h l o r i n e C o n t e n t D e t e r m i n a t i o n " , (See T a b l e No» 5)a w e a r s q u o t i n g r a t h e r f u l l y f r o m t h e pub-l i c a t i o n , a s f o l l o w s : "The o r g a n i s m S. a g a l a c t i a e * h a s a v e r y d e f i n i t e g r o w t h c h a r a c t e r i s t i c when grown as p u r e c u l t u r e i n i n f u s i o n b r o t h ; 1, e. , h e a v y g r o w t h i n t h e b o t t o m and. up t h e s i d e o f t h e t e s t t u b e . I n w o r k i n g w i t h m i l k s a m p l e s t h e s e n i o r a u t h o r n o t e d t h a t when some s a m p l e s w ere i n c u b a t e d , i n s m a l l - b o r e g l a s s t u b e s f o r a . p e r i o d o f t w e n t y - f o u r h o u r s t h e r e w o u l d a p p e a r on t h e s i d e s m a l l r o u n d f l a k e s w h i c h were s e e m i n g l y d e n s e r t h a n t h e s u r r o u n d i n g medium. H a v i n g n o t e d t h e c h a r a c t e r i s t i c groxvth. i n i n f u s i o n b r o t h and k n o w i n g t h a t m i l k i s a n a t u r a l h a b i t a t o f the o r g a n i s m , he e x a m i n e d t h i s c o l o n y l i k e g r o w t h i n m i l k * A l s o r e f e r r e d t o a s S t r e p t o c o c c u s M a s t i t i d i s . 19. and f o u n d i t t o be S. a g a l a c t i a e . However, s u c h . g r o w t h was somewhat hard, t o s e e , so means were s o u g h t by w h i c h t h e g r o w t h c o u l d be r e c o g n i z e d more e a s i l y , " K l i m m e r * , i n h i s serum a l k a l i a l b u m i n a t e s a c c -h a r o s e a,gar, a d d e d a s a t u r a t e d aqueous s o l u t i o n o f brom-c r e s o l - p u r p l e . He s t a t e d t h a t on t h i s a g a r S. a g a l a c t i a e c o l o n i e s i n t w e n t y - f o u r h o u r s a r e a d a r k y e l l o w s u r r o u n d e d by a s m a l l c l o u d y b o r d e r and a l a r g e r b r i g h t y e l l o w h a l o . B a k e r and Van S l y k e * had shown t h a t b r o m - c r e s o l - p u r p l e when a d d e d t o m i l k p o s s e s s e s p r o p e r t i e s w h i c h make i t a n i n d i c a t o r f o r t h e c o l o r i m e t r i c d e t e r m i n a t i o n o f h y d r o g e n - i o n c o n c e n t r a -t i o n I n m i l k . F rom t h e s e o b s e r v a t i o n s , t h e s e n i o r a u t h o r c o n c l u d e d t h a t a change i n c o l o r a f t e r t h e a d d i t i o n o f t h i s dye t o t h e m i l k would, i n d i c a t e a c i d p r o d u c t i o n and p o s s i b l y p e r m i t e a s i e r r e c o g n i t i o n o f t h e f l a k e s , t h u s f a c i l i t a t i n g t h e i d e n t i f i c a t i o n o f S • a g a l a c t i a e . "The H o t i s t e s t , t h e r e f o r e , c o n s i s t s i n a d d i n g 0.5" c c , o f a s t e r i l e 0.5 p e r c e n t aqueous s o l u t i o n o f b r o m o - c r e s o l -p u r p l e ( d i b r o m o r t h o c r e s o l s u l p h o n p h t h a l e i n ) t o 9*5 cc,. o f m i l k p r e v i o u s l y m e asured i n t o a s t e r i l e t e s t t u b e . A f t e r t h e t u b e i s i n v e r t e d s e v e r a l t i m e s t o m i x t h e c o n t e n t s , t h e sample i s . i n c u b a t e d a t 37*5° •^• f o r t w e n t y - f o u r h o u r s and t h e r e s u l t s a r e t h e n o b s e r v e d . A m i x e d sample o f m i l k , c o n s i s t i n g o f an e q u a l - q u a n t i t y f r o m e a c h q u a r t e r o f t h e u d d e r , o r s a m p l e s f r o m * T h e s e r e f e r e n c e s a r e g i v e n i n t h e b i b l i o g r a p h y o f t h e p u b -l i c a t i o n (6) f r o m w h i c h t h i s q u o t a . t i o n i s ma.de.-i n d i v i d u a l q u a r t e r s may be u s e d f o r t h e t e s t . However, t h e i n d i v i d u a l q u a r t e r sample i s p r o b a b l y more a c c u r a t e , e s p e c i a l l y when s t r e p t o c o c c i a r e p r e s e n t i n s m a l l numbers i n o n l y one q u a r t e r . " I m m e d i a t e l y a f t e r t h e m i l k and i n d i c a t o r have been m i x e d , a deep p u r p l e c o l o r r e s u l t s . The c o l o r i s due t o t h e f a c t t h a t t h e a c i d i t y o f m i l k (pH 6 . 3 o r o v e r ) i s l o w and t h e r e f o r e i n t h e u p p e r r a n g e o f t h e i n d i c a t o r (pH y.2,,yellow t o pH 6 . 8 , p u r p l e ) . I f s t r e p t o c o c c i a r e p r e s e n t , t h e c o l o r c h a n g e s f r o m p u r p l e t o a y e l l o w shade d u r i n g i n c u b a t i o n as a r e s u l t o f t h e p r o d u c t i o n o f a c i d f r o m l a c t o s e by t h e s e o r g a n i s m s . I n a d d i t i o n t o t h i s c h a n g e , i f S. a g a l a c t i a e i s p r e s e n t a s m a l l f l a k e s o r b a l l s o f g r o w t h , f r o m 0 , 5 t o 4 mm. i n d i a m e t e r , u s -u a l l y f o r m on t h e s i d e o f t h e t u b e . These g r o w t h a r e a p p a r e n t l composed o f m i l k c u r d and e n t w i n e d c h a i n s o f S. a g a l a c t i a e , w h i c h i s t h e p r e d o m i n a n t s p e c i e s f o u n d i n s t r e p t o c o c c i c mas-t i t i s . T h e i r o c c u r r e n c e has b e e n o b s e r v e d f r e q u e n t l y i n i n -c u b a t e d m i l k t o w h i c h no i n d i c a t o r h a s been a d d e d , b u t i n t h i s t e s t t h e y a r e s e e n more r e a d i l y b e c a u s e , a s a r u l e , t h e y a p p e a r a s c a n a r y - y e l l o w s p o t s a t t a c h e d t o t h e s i d e o f t h e t u b e and i m -m e d i a t e l y s u r r o u n d e d b y a h a l o o f t h e same c o l o r , w h i c h i n t u r n g r a d u a l l y merges w i t h t h e d a r k e r b a c k g r o u n d o f t h e colum n o f m i l k , The number o f f l a k e s v a r i e s f r o m s e v e r a l t o c o u n t -l e s s and t h e s i z e f r o m p i n p o i n t , when t h e r e a r e many f l a k e s , t o a b o u t 4 mm. i n d i a m e t e r when o n l y a few a r e p r e s e n t . When S. a g a l a c t i a e i s p r e s e n t i n t h e s a m p l e , i t i s r a r e t h a t no f l a k e s o r c l u m p s a r e n o t e d * 2.1, " V a r i o u s s h a d e s o f c o l o r a r e e n c o u n t e r e d i n t e s t s w h i c h may be c o n s i d e r e d s u s p i c i o u s o r p o s i t i v e . These r a n g e f r o m a b l u e g r a y o r a d i s t i n c t o l i v e c o l o r t h r o u g h y e l l o w i s h g r e e n and g r e e n i s h y e l l o w t o a p r o n o u n c e d y e l l o w o r g o l d e n hue. When t h e f i r s t m e n t i o n e d c o l o r s o c c u r , an- a d d i t i o n a l t w e n t y - f o u r h o u r s ' i n c u b a t i o n w i l l u s u a l l y b r i n g o u t t h e y e l -l o w c o l o r . However, i f t h e i n d i s t i n c t s h a d e s a r e a s s o c i a t e d w i t h t h e o c c u r r e n c e o f f l a k e s on t h e s i d e o f t h e t u b e s , w h i c h i s v e r y o f t e n t h e e a s e , no f u r t h e r i n c u b a t i o n i s n e c e s s a r y t o i n d i c a t e a p o s i t i v e r e s u l t , A l i t t l e e x p e r i e n c e w i t h t h e t e s t w i l l e n a b l e t h e o p e r a t o r t o r e c o g n i z e t h e s e p o i n t s w i t h o u t d i f f i c u l t y , . " D u r i n g t h e c o u r s e o f t h i s w o r k 753 s a m p l e s o f m i l k were s u b j e c t e d t o t h e H o t i s t e s t f o r t h e d e t e c t i o n o f m a s t i t i s s t r e p t o c o c c i and a l s o t o t h e b l o o d - a g a r p l a t e method o f d e t e r -m i n i n g t h e p r e s e n c e o f t h e s e o r g a n i s m s . The m a j o r i t y o f t h e s e s a m p l e s were f r o m i n d i v i d u a l q u a r t e r s and t h e r e m a i n d e r were' c o m p o s i t e s a m p l e s t a k e n d i r e c t l y f r o m t h e f o u r q u a r t e r s o f t h e u d d e r , " P o s i t i v e r e a c t i o n s t o t h e H o t i s t e s t , s h o w i n g b o t h t h e t y p i c a l c o l o r change and t h e f l a k e s on the s i d e o f t h e t u b e , o c c u r r e d i n ^60 s a m p l e s i n w h i c h t h e b l o o d - a g a r p l a t e s showed m a s t i t i s - s t r e p t o c o c c i . P o s i t i v e c o l o r r e a c t i o n s w i t h o u t t h e c h a r a c t e r i s t i c f l a k e s o c c u r r e d i n 1 0 8 a d d i t i o n a l s a m p l e s i n w h i c h t h e b l o o d - a g a r p l a t e s showed s t r e p t o c o c c i , a nd i n 2 5 i n w h i c h no s t r e p t o c o c c i were f o u n d . N e g a t i v e r e a c t i o n s t o t h e 2 2 . H o t i s t e s t o c c u r r e d i n 60 s a m p l e s , o n l y 13 o f w h i c h showed s t r e p t o c o c c i on b l o o d - a g a r p l a t e s . "A t o t a l o f 668 samples were c l a s s i f i e d p o s i t i v e b'oth by t h e H o t i s t e s t a,nd by t h e b l o o d - a g a r method, and 47 w e r e c l a s s i f i e d a s n e g a t i v e . I n o t h e r words t h e two methods were i n p e r f e c t a g r e e m e n t f o r 71?» o r 95 p e r c e n t , o f t h e 753 s a m p l e s t e s t e d . . T h i s f i g u r e may be c o n s i d e r e d an a p p r o x i m a t e measure o f t h e a c c u r a c y o f t h e H o t i s t e s t , i f t h e r e s u l t s i n -d i c a t e d by t h e b l o o d - a g a r p l a t e s a r e assumed t o be 100 p e r c e n t a c c u r a t e . " I t i s n o t p o s s i b l e a t t h i s t i m e t o o f f e r a d e f i n i t e e x p l a n a t i o n f o r t h e v a r i a t i o n i n c o l o r s , o r f o r t h e a b s e n c e o f f l a k e s i n some o f t h e p o s i t i v e t e s t s . W h i l e t h e H o t i s method was b e i n g d e v e l o p e d , b l o o d - a g a r p l a t e s were made-in o r d e r t o d e t e r m i n e t h e p r e s e n c e and t h e number o f s t r e p t o c o c c i i n t h e m i l k s a m p l e s , T h e r e was some c o r r e l a t i o n between t h e number o f o r g a n i s m s a n d t h e c o l o r o f t h e sample as w e l l a s t h e number o f f l a k e s , b u t i t was n o t s i f f i c i e n t l y c l o s e t o e x p l a i n e n t i r -e l y t h e v a r i a t i o n t h a t o c c u r r e d . T h e r e i s some p o s s i b i l i t y t h a t a n i n v e s t i g a t i o n o f t h e l a c t o s e c o n t e n t , pH v a l u e , and change i n o t h e r c o n s t i t u e n t s o f t h e s a m p l e , t o g e t h e r w i t h t h e b a c t e r i a l c o u n t , m i g h t y i e l d t h e i n f o r m a t i o n . However, s i n c e t h e method was d e s i g n e d p r i m a r i l y to d e t e c t t h e p r e s e n c e o f S. a g a l a c t i a e i n m i l k , v e r y l i t t l e w o r k has been done t o e x p l a i n t h e s e v a r i a t i o n s . 23. " I n o r d e r t o o b t a i n c l e a r - c u t r e s u l t s w i t h t h e t e s t i t i s h i g h l y d e s i r a b l e t o c o l l e c t t h e m i l k d i r e c t l y f r o m t h e a n i m a l and a s a s e p t i c a l l y a s p o s s i b l e . When t h e s ample i s t a k e n f r o m t h e p a i l o r i n a manner t o p e r m i t g r o s s c o n t a m i n -a t i o n w i t h r o d s j m i c r o c o c c i , and s a p r o p h y t i c s t r e p t o c o c c i , a p o s i t i v e r e a c t i o n may be masked by p r o t e o l y s i s o r a l k a l i f o r -m a t i o n s o r b o t h , and o c c a s i o n a l l y a f a l s e p o s i t i v e r e s u l t may ensue f r o m c o n t a m i n a t i o n w i t h l a c t i c - a c i d b a c t e r i a . , A l l t h e s e c o n d i t i o n s have been e n c o u n t e r e d i n t h e c o u r s e o f t h i s w o r k . The method a s c o n s t i t u t e d a t p r e s e n t , t h e r e f o r e , i s n o t s u i t a b l e f o r t h e e x a m i n a t i o n o f m i l k f r o m c a n s o r a t r e c e i v -i n g ' s t a t i o n s * " A d v a n t a g e s o f t h i s t e s t a r e ; (1) O n l y a s m a l l am-o u n t o f e q u i p m e n t i s r e q u i r e d , s u c h a s a s u p p l y o f s t e r i l e t e s t tubes- 12 mm, i n d i a m e t e r , r a c k s f o r t u b e s , s t e r i l e brom-c r e s o l - p u r p l e i n d i c a t o r , a s t e r i l e p i p e t t e f o r a d d i n g t h e i n -d i c a t o r , a n d a n i n c u b a t o r s e t a t 37* 5° 0; (2) t h e m i l k sample ha.ving been c o l l e c t e d i n a s t e r i l e t u b e o r b o t t l e , i t may be p o u r e d i n t o t h e t e s t t u b e g r a d u a t e d f r o m 9 * 5 c c . , t h u s ob-v i a t i n g t h e u s e o f a p i p e t t e f o r e a c h sample and i n c r e a s i n g t h e number o f t e s t s t h a t c a n be s e t up i n a g i v e n t i m e ? a n d (3) t h e m i l k s e r v e s a s t h e c u l t u r e medium and i n a s m u c h a s t h e b r o m - c r e s o l - p u r p l e i s a p p a r e n t l y ' b a c t e r i o s t a t i c f o r some o f t h e u d d e r m i c r o c o c c i , a t l e a s t f r o t h e f i r s t t w e n t y - f o u r h o u r s , v e r y l i t t l e i n a c c u r a c y i s e x p e r i e n c e d , f r o m t h i s o u r c e when p r o p e r p r e c a u t i o n s have been o b s e r v e d i n c o l l e c t i n g t h e sam-p l e , 24. Summary "A s i m p l e method f o r t h e d e t e c t i o n o f m a s t i t i s s t r e p t o c o c c i i n m i l k t h a t c a n be r e a d i l y u s e d on l a r g e num-b e r s o f s a m p l e s and t h a t r e q u i r e s but l i t t l e e q u i p m e n t , i s d e s c r i b e d . T h i s t e s t i s d e s i g n a t e d a s t h e H o t i s t e s t i n r e c o g n i t i o n o f t h e work o f t h e s e n i o r a u t h o r . , "The method c o n s i s t s i n a d d i n g 0 * 5 c c » o f a s t e r i l e 0 . 5 p e r c e n t a q u e o u s s o l u t i o n o f brom-ere s o l - p u r p l e t o 9 * 5 c c , o f m i l k c a r e f u l l y c o l l e c t e d d i r e c t l y f r o m t h e a n i m a l . A f t e r t h e s a mple i s mixed, i t i s i n c u b a t e d f o r t w e n t y - f o u r h o u r s a t 3 7 - 5 ° C. and t h e r e s u l t s a r e o b s e r v e d , "A c h a r a c t e r i s t i c change i n t h e c o l o r o f t h e sample a f t e r i n c u b a t i o n t o g e t h e r w i t h t h e o c c u r r e n c e o f f l a k e s o r b a l l s o f growth i n d i c a t e s t h e p r e s e n c e o f S. a g a l a c t i a e „ " E x a m i n a t i o n o f 7 5 3 s a m p l e s o f m i l k by t h e H o t i s t e s t and. by t h e b l o o d - a g a r method showed t h e two t e s t s t o be i n p e r f e c t a g r e e m e n t f o r 7 1 5 ? or 9 5 p e r c e n t , o f t h e s a m p l e s . T h i s f i g u r e may be c o n s i d e r e d a n a p p r o x i m a t e measure o f t h e a c c u r a c y o f t h e H o t i s t e s t , i f t h e r e s u l t s i n d i c a t e d by t h e b l o o d - a g a r p l a t e s a r e assumed t o be 1 0 0 p e r c e n t a c c u r a t e . " P r o c e d u r e f o l l o w e d I n C l a s s i f y i n g Q u a r t e r s Tfith R e g a r d to M a s t i t i s I n f e c t i o n ; The w o r k e r s i n v e s t i g a t i n g m a s t i t i s a r e i n g e n e r a l a g r e e m e n t t h a t t h e v a r i o u s t e s t s used, f o r d e t e c t i n g m a s t i t i s do n o t n e c e s s a r i l y y i e l d s t r i c t l y c o m p a r a b l e r e s u l t s * C e r t a i n 2 5 -t e s t s , f o r i n s t a n c e D i s l a c , a r e a p p l i c a b l e f o r use by d a i r y -men, w h i l e o t h e r s may be employed by v e t e r i n a r i a n s o r t e c h n i -c a l l y t r a i n e d f i e l d w o r k e r s , a n d s t i l l o t h e r s c a n be us e d o n l y u n d e r l a b o r a t o r y c o n d i t i o n s , The v a r i o u s t e s t s v a r y i n t h e i r s e n s i t i v e n e s s . One t h a t d e t e c t s a n a d v a n c e d s t a g e o f i n f e c -t i o n may be o f l i t t l e o r no use i n d e t e c t i n g t h e m i l d e r c a s e s o f m a s t i t i s * The o p i n i o n e x p r e s s e d by R o g e r s (3) i n d i s c u s -s i n g t h e a p p l i c a t i o n o f a number o f t h e more w i d e l y used t e s t e i s " p o s i t i v e a c t i o n s w i t h a n y two o f t h e s e t e s t s a r e u s u a l l y c o n s i d e r e d c o n c l u s i v e e v i d e n c e o f t h e p r e s e n c e o f t h e d i s e a s e . The number o f p o s i t i v e l y r e a c t i n g cows f o u n d w i l l be d i r e c t l y p r o p o r t i o n a l t o t h e number o f t e s t s e m p l o y e d " . T h i s o p i n i o n a p p e a r s t o be h e l d by most i n v e s t i g a t o r s * . I n t h i s i n v e s t i g a t i o n t h e a u t h o r s have c l a s s i f i e d q u a r t e r s I n t o P o s i t i v e , D o u b t f u l * and N e g a t i v e g r o u p s r e mas-t i t i s i n f e c t i o n a c c o r d i n g t o t h e f o l l o w i n g methods? P o s i t i v e Group £ ( l ) When two o r more t e s t s i n d i c a t e t h e p r e s e n c e o f a m a s t i t i s i n f e c t i o n , Or ( 2 ) When one t e s t i s p o s i t i v e and two o r more a.re n e g a t i v e , *The word " d o u b t f u l " and t h e l e t t e r "D" a r e used t h r o u g h o u t t h i s t h e s i s t o i n d i c a t e a d e p a r t u r e f r o m t h e n o r m a l , w h i c h i s n o t c o n s i d e r e d s u f f i c i e n t l y d e f i n i t e i n c h a r a c t e r t o j u s t i f y a " p o s i t i v e " i n t e r p r e t a t i o n . 26\. o r (3) When t h e H o t i s r e a d i n g i s P 4 - ( i n d i c a t i n g t h a t c h a r a c -t e r i s t i c c o l o n i e s o r f l a k e s a p p e a r e d on t h e s i d e s o f t h e t u b e i n t h e H o t i s T e s t , - D o u b t f u l G roup? ( l ) When one, only,, t e s t i n d i c a t e s t h e p r e s e n c e o f a mas-t i t i s i n f e c t i o n ( e x c e p t t h e " H o t i s p+" ) f o r (2) When two o r t h r e e t e s t s g i v e r e s u l t s r e q u i r i n g a " d o u b t f u l . " i n t e r p r e t a t i o n * N e g a t i v e G r o up : ( l ) When a l l t e s t s a r e i n t e r p r e t e d a s i n d i c a t i n g t h e a b-s e n c e o f m a s t i t i s , o r ( 2 ) When one t e s t i s g i v e n a " d o u b t f u l " i n t e r p r e t a t i o n and t h e r e s t a r e n e g a t i v e . PRESENTATION OP RESULTS AND PIS CUSS ION. E x p l a n a t i o n s R e l a t i v e t o P r e s e n t a t i o n o f M a t e r i a l t R e s u l t s o f t h e v a r i o u s t e s t s employed, a r e c o m p i l e d I n T a b l e s No. 1, No. 2 a n d No* 3s and a c o n c l u s i o n r e g a r d i n g m a s t i t i s i n f e c t i o n i s p r e s e n t e d f o r e a c h q u a r t e r o f ea.ch cow's u d d e r by t h e method a l r e a d y o u t l i n e d . I n T a b l e No. 4, c a s e h i s t o r i e s r e l a t i v e t o t h e ex-p e r i m e n t a l a n i m a l s a r e l i n k e d w i t h t h e c o n c l u s i o n s r e m a s t i t i s I n f e c t i o n a r r i v e d a t i n T a b l e s No. 1, No. 2 and No. 3 * T a b l e No, 4, t h e r e f o r e , i s l a r g e l y a d i s c u s s i o n p r e s e n t e d i n t a b l e f o r m . I n T a b l e No. 5» r e s u l t s o f t h e c h l o r i n e and H o t i s t e s t s , a r e c o m p a r e d . T h i s c o m p a r i s o n i s "based on r e s u l t s o f , t h e s e two t e s t s where b o t h were u s e d ->- t h a t i s , i n T a b l e s No. 2 and No. 3• The c o r r e l a t i o n o f p o s i t i v e r e s u l t s i n T a b l e s No, 1, No. 2 and No„ 3s 1 S i n d i c a t e d by t h e use o f r e d t y p e . I n t h e " q u a r t e r " c o l u m n , r e d t y p e i s used when a q u a r t e r o r t e a t has been i n j u r e d , o r when abnormal m i l k has been p r o d u c e d a t some t i m e , I n t h e " b a c t e r i a l c o n t e n t " c o l u m n , r e d means t h a t a c i d p r o d u c i n g b a c t e r i a were p r e s e n t i n t h e " p l a t e " test» E x p l a n a t i o n s o f l e t t e r s a n d A b b r e v i a t i o n s . U s e d i n T a b l e s ; I n t h e " Q u a r t e r " Columnt K E -~ R i g h t f r o n t , L E L e f t f r o n t * RE — R i g h t hind„ LPT •— L e f t h i n d * I n t h e " D i s l a c " Column t N -~ N e g a t i v e ( l i g h t s l a t e ) * S I . 1 — S l i g h t l y p o s i t i v e ( d a r k s l a t e ) P -- P o s i t i v e P4~ — V e r y p o s i t i v e (deep p u r p l e } * . I n t h e " H o t i s T e s t " Column % N — N e g a t i v e ( n o r m a l blue)» D — D o u b t f u l ( c o l o r change b u t n o t c a n a r y y e l l o w ) . P — P o s i t i v e ( c a n a r y y e l l o w ) P+ — P o s i t i v e ( a n y c o l o r w i t h f l a k e s o r c o l o n i e s ) * Alk.. -- A l k a l i n e (deep b l u e o r deep p u r p l e ) . 28. I n t h e " H o t i s T e s t " Column — C o n t i n u e d . P r o t e a . •— P r o t e o l y s i s ( p a r t l y d i g e s t e d ) . Ga.s — P r e s e n c e o f g a s . I n t h e " C o n c l u s i o n r e M a s t i t i s I n f e c t i o n " Columns. F —• N e g a t i v e . D Doubtful» P -~ P o s i t i v e * 29 c H I r H to — * c pa o CQ EH •I-l P» b=- J_-c o o o o ft fl o < ! A•rH CQ ft c: m m •H - P ft O • H S <J - P <£, PQ cc CC CC « g E fl <! a. H u ft fl 6 3 Hi c S ft •rH 1 En O CQ feS O O O c o I c fl Fl o M H < C M fl H <H CO > EH H ft fl O o CC EH EH 8* O CO ft EH , f l < ! - H fl fiw E H <! CQ | . C cC CQ o O O fe - H 0 fl fl . H - P CH O O CQ 0 * H fl ' H 3 M + i M - P rH • -p 02 fl rH 0 fl -p fl C O o o • o H o C6 • H rH • ?H 0 CQ fl.' ft f-l - P fl O a. «Hr pq OJ -p fl 0 - P fl o o I CQ H fi O CC5 rH o o o o r-i fl fl • H O O r i o o O fe rH ' x H |2i FH • fl OJ O osfe 3 <: ft o o a H CQ •H rH I fH • 3 0 a?-P CQ i o t2S I ft s p aj cc cc 0 - P CC <+H o 1 • ft hO g fl CO - H CC r H o o O fl fl ft iz, fl ft ft fl ft p f t f t f t |ZJ ft p ft 1 1 O O O O O O O O CO C^CTsrO CM C-H o o o o o o o o o o H C T N O H O N • CM CMMD "si-H n o > no I 1 M3 >- CMMD r-\ r-{ r-i r~\ • • • • tr\ r o ^ O O O O I CM CM CO CA vO r l H H H H C M M 3 • • • • O O H CM iH O O o VO ON O "sh CJ \ I> -v O H t>-C \ i I C N O Y C O H H H H CM -Hr H/ m o • • • • <r ifNtrwo ft ft ft ft ft ft ft ft • • • • • « + H ft H H r H r H rH r H i ~ ft ft ft CQ cccc ra to ra to cc ft ft w w PH fl PH fl r H • f> C O O O N fl r-i ft ft fl fl ft fl fl ro \ r-i CM OJ • \ 0 i> ro O O N fl H ft ft fl fl rn|pj CO r H ro O O fl H CM fl fl o o O N ro H/ ro r H r H CO ro ro r o C ^ v O v O H H r H H O N O V O V O • « » • ft ft H fl H PH CC CC ft K W fl PH fl ro CM CO CM CM • N O > ro O O -rS) r—l 30 o C o o CQ I in o a - H <D - P ft CD -H C M -P M ft ft PH ft ft PH PH PH fx, p< p< p< o ft P^  ft ft ft ft ft -p 0 CD -P •H +5 o o O o CD a 1 •H rl H O • i—1 ft O o | !==! CD H £1 o3 H CQ rl a o o3 H M ft c i o •H O • M O O H C i O H •rl ft 3 ,I.J> CQ h O O o o o O CD M O O o o o O -P 1 o- CM n o o o ft c OO ft ft o OO O -o • o H ft o cfl •H rl • rl CD CQ a- ft U •p ft o o 1 o 1 i 1 1 t 03 CM pq CM o O OOrH o o o • f t rl O X fai) (D ft <i ft o o o3 ft CQ •H n rl • 03 U CQ £ -S ft 03 • CD O H -P ft ft o3 ca I ft bD| S C 03 - r l | CD ft Ml -p a s CO ^ 05 -H ft O CQ ft £ • o o O ft o o o o I CO "3" ft ft *# O -I I I o o CM CM I I I CM PO O -iov ON m CM O O CM r H CM CM CM CM CO O N r o o • • • • ft O N C M v O CM O-MD l r \ O C O ft o CM ft CM CM C N - OO ft CO • • • • O - CO CM CO r O l N O O O - O - O - O O ft ft ft ft O O C O ft • • • • ftftftft ftftftft p< FH ft ft PR ft ft ft ft ft ft ft 1 r \ ft • v O $> OO O C A N ft no fx. ft Hi ft c4 ft PI ft OO s . CM CM CM • v O t> o o ,0 O N ft ft ft ft f3q £r} JX; ft ft ft CO i> o o O OA N ft v O P O O - C M VO O - o O O v O i N - O - O -ft ft ft ft O - O OVX> ft ftftftft CQ ft ft ft 1X! ftftftft OO X CM CO CM CM • \ D i> o o C ON ft" ft O O O O O O cM *t i r \ * at M « c o o n i r \ ft o O I CA CM CO O O - O N O iS-oo oco co ft CM ft ft P O O ft o n • • * • ftftftft ftftftft ftftftft CO ft • v O t> OO O C A ft ft 1r \ 31. 0 a •H -P g ft O ft CD r H ft CO EH rH O S3 o o I CO I CO, 02 O • •H CD •rl -P CH CQ CD -rl Ej ^ U -P M +i -P a - p e o o ft CO •rl rl 0 -P o CO ft CQ rl ft 00 -P 0 • p fl c o 0 C •rl rl O r H x: l CQ rl S o CO r H rH ft C l O c o •H O • ft 0 O r H a ° td O O r H O r O r H ft O O m ' H ft rl • . . o \ bo 0• o Oi f t tft <ij ft o c co r H CQ •H ft I U •• CO rl 3 0 Of -P CQ S CQ • 0 -P O rl CO CO ft ft CO I • ft bX 0 ft id - p s a CO ft <0 -H ft O CO ft ftftftft ft ft P ft ft ft ft ftft ftP ftP^O o o o o o o o o oo o-if\co ft ft a-' • ft CQ o fH o ft o "3- o OJ ft O ft CA-CM ^ C O CM C A O ^ t H O CM CM CM CM v O O O C A • « • » v O O - v O Vr\ + ft ft ft ft ftftftft ftftftft H ^ CO CM C M V O . no > C A O H o o o o o o o o ft OO I f N v O CM r O ft ft <tf o o o o o o o o ft 00 IrwO * \ *\. ** CM i - O H ^j-CM r o t o o o o CM ir\co CM CM ft ft C A c O l T N C M . . . . ftftftft co ft P R ft ft CO Irs ftvO • C O l> C A o o o o o o o o C A C M C A C O CM O CM CM O O O O C A C M CM O I CM CM CO o o C O CO •sj- irwo o-ftCOvO C O C A C A v O O -ft ft ft ft ^ - V O O - C A • • # • + ftftftft ft ft ft ft ft ft ft ft -4* oo CM C M v O • OO i> C A O ft ft o o o o o o o o C A C O ft O ft CM OO o o O C A ft CM v O O O O O O l r \ t A - \ Q O -ft ft ft ft >Hr C A v O C O . . . . f^- sf- ^ ftftftft • . • . i—I i—I i—j i—I CO CO CQ CO ft ftftftft ft . f t f t f t f t O O O O O O CM <H- CM CM ft oo^f- oo\r\ l T\CM l T \ C A Xr\ ^  to ftftftft ft OO ftlT\ • • • • •d- ^  ^ t^-ft ft m ft • ft co ft ft ft ft ft ft oo Irs ftvO • on *> o^ O rH OO CM CMM3 • on > O N O ft !zs o o l O ftj CA 32. i CQ I C CC CQ O O fl -r-i 0 -P C H o O CQ CD -H fl -H O 3 fn -P rH -P • •p CQ fl u m fl -p fl 00 o o • o 1—1 o aj •H u • fH 0 CQ (D ft u •P ft O cC <=* m cv +3| fl CD •P fl I fl o CQ M -H O • S O fl, CC rH CD O r-i p, ft c o -Hi c ci o o 0)1 fll • fl • H O O rH M 1 O rOrH .H O O O fe H i • rH fl M • fll 0 \ W CD O O l f l fl <| ft O o 05 H CQ •H fl I rH CC fH 3 CD <3?-P -P cC CO ft taOj 6 fl CC "H H I <D ft bo -p a A crj ft cC -H fl O CO H fl fl fl fl flflflfl fl fl pn fl ft ft fl, fl fl fl ^ C O O N O N O N O - O N O W H O O rH rH rH rH O- <dr CM CM n o n o r o r o O N O N < t N O O N O N r o O O O r-i H H i—I i—I r o r h r o ^ \r\CG no O N «?+- pr ; CM C\J H rH CM rH H CM CM r o H O N r o C ^ . . . . . . . . n o r o r o r o m \ o c n ONtr\ O N o O N CXI CM Vf\ r o ONNO l r \ CO H ' H - IJTN ^ j - 1 r \ C M O N ^ r o o c M CO rH CM rH r H r H ^ t - < H -O-lfN^J- \ T N C M O - H r O ON 'H- VO 1_CN <H r o rH CM H fl flflflfl flflflfl + flflflfl ft + ft fl * ft i—I i—I co co flflflfl flflflfl no CM r o CM • N O O r o CD O N fl rH ft ft fl fl ft ft fl ft ft ft fl fl ri fl 3 A no rH no CM • N O O no CD ON fl rH CM O -CM •NO O r o CD O N fl H flflflfl flflflfl to o-CM •NO O r o CD O N fl rH ft ft-r + • • ft fl H H CO CO flflflfl fl fl fl ft fl CD — -CD fl iM O fl -H -P -P o fn 0 CD ft -P -H O •P CD CQ fl cC m ft cC • fH O CD -P -P r o M 0 L T \ cC 3 H a1 fl M H fl -H •H S fl O 0 H -H -P • -P 0 <vh o f-i fl o H 0 ft CQ 0 O G -P o >> 0 rH CQ fl CC O 0 o ft CQ aj 0 aj fl r o f t fl H CQ fH 0 CM • N O O r o 0 O N fl rH o fl c fH cd - 3 H O D H CM • NO O r o 0 O N fl H & • o o o fl C O O N o r-l CM r o 33. 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CD to +3 43 to to to bo >ci ft CD rH 43 CQ O to to to to totototo toto toto totototo totototo toto toto-to to to ft ft ft ft H CM 5 ft ft to ft ft r o 57. - ; S U I H E Y 0? TABLS NO_4_ The i n f o r m a t i o n p r e s e n t e d i n T a b l e No. 4 i s most r e a d i l y and c l e a r l y s u m m a r i z e d by g r o u p i n g ( l ) cows, and (2) . q u a r t e r s 5 on t h e b a s i s o f "Case H i s t o r y " a n d c l a s s i f y i n g them by numbers and p e r c e n t a g e s i n t o p o s i t i v e , d o u b t f u l and nega -t i v e c o n c l u s i o n s r e m a s t i t i s i n f e c t i o n . When a r e p e a t t e s t on a cow r e s u l t e d i n a d i f f e r e n t c o n c l u s i o n r e m a s t i t i s i n f e c t i o n t o t h a t o f t h e f i r s t t e s t , t h a t cow h a s been r e p r e s e n t e d a s "V1 i n e a c h o f two " c o n c l u -s i o n " classifications.» T h i s method h a s been u s e d a l s o i n c l a s s i f y i n g t h e q u a r t e r s t e s t e d . . I t i s seen t h a t a l l q u a r t e r s i n Group 1 a r e n o r m a l . I n G r o u p s 2, 3» and 4, h o w e v e r , t h e r e a r e b o t h n o r m a l a n d a b n o r m a l q u a r t e r s . I n t h e s e l a s t t h r e e g r o u p s , t h e r e f o r e ; more«than one c l a s s i f i c a t i o n h a s been f o u n d n e c e s s a r y i n o r d e r t o show c l e a r l y t h e r e l a t i o n s h i p e x i s t i n g between " c a s e h i s t o r y " and t h e i n c i d e n c e o f m a s t X t i s . Group 1. A p p a r e n t l y n o r m a l cows (no i n j u r i e s and no p h y s i -c a l l y a b n o r m a l m i l k p r o d u c e d a t a n y t i m e , i n c l u d i n g t i m e o f s a m p l i n g ) . T h e r e a r e e i g h t (8) cows i n t h i s g r o u p : -N o s , 7, 1?, 16, 21, 25? 51* 5 2 an<3 53 • On a "Cow Basis", P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n 3"! o r 43.7%• D o u b t f u l 11 " " " — 3 o r 3 7 . 5 f . N e g a t i v e " " " " l i o r 18.8,3. 58. Group 1 C o n t i n u e d On a " Q u a r t e r Basis.".. T h i r t y - t w o (32) a p p a r e n t l y n o r m a l q u a r t e r s (no ' i n j u r i e s and no p h y s i c a l l y a b n o r m a l m i l k p r o d u c e d a t any t i m e , i n c l u d i n g t i m e o f s a m p l i n g ) . P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n 5it o r 17 .2$, D o u b t f u l " " " " — 13£ o r 4 2 . 2 ^ . N e g a t i v e ' " " " " -- 13 o r 40.61, 6roujQ^_2» Cows w i t h i n j u r i e s t o u d d e r o r t e a t s b u t w h i c h have n o t been known t o p r o d u c e o t h e r t h a n a p p a r e n t l y n o r m a l m i l k . T h e r e a r e t w e n t y - o n e (21 ) cows i n t h i s g r o u p N o s . 8 5 14, 18,. 2o, 22, 23, 29» 30 5 31s 3.2? 33s 3'8, 3.9*. 4 0 s 4 2 , 4 3 , 44, 45 , 46 s 47, and 48,. On_a__"_Cpw J B a s i s " . P o s i t i v e C o n c l u s i o n s r e I v l a s t . i t i s I n f e c t i o n — 1J? o r 71.4^, D o u b t f u l " " " " — 5 o r 23.8:^. N e g a t i v e " " " " 1 o r 4 J On a "Q.uarter B a s i s " . . S i x t y - o n e ( 6 l ) a p p a r e n t l y n o r m a l q u a r t e r s (no i n -j u r i e s and no p h y s i c a l l y a b n o r m a l m i l k p r o d u c e d a t any time , i n c l u d i n g t i m e o f s a m p l i n g ) . P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n --• 14-£- o r 23 ,Q%<. D o u b t f u l " " »• " — 14£ o r 23.8^. N e g a t i v e " " » « -- 32 o r 5 2 .4$. 59. Q£ o up_ 2 - Con t i n u e c t T w e n t y - t h r e e (23) q u a r t e r s w h i c h have had i n j u r i e s b u t w h i c h h a v e n o t been known t o p r o d u c e o t h e r t h a n a p p a r e n t l y ' n o r m a l m i l k . P o s i t i v e C o n c l u s i o n s r e l a s t i t i s I n f e c t i o n 9 o r 39.I/L D o u b t f u l " " " " — 5 o r 21.8?$. N e g a t i v e " 11 " " — y o r 3.9,1%* G r o u p 3« Cows w i t h a m a s t i t i s h i s t o r y ( h a v i n g a t some t i m e p r o d u c e d p h y s i c a l l y a b n o r m a l m i l k ) . . b u t w h i c h a t t i m e o f s a m p l i n g were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k f r o m a l l f o u r q u a r t e r s . T h e r e a r e f i f t e e n (15) cows i n t h i s g r o u p s -Nos.: 1, 2. 3, 4,. 9. i o , 13, 17, 19, 24s. 26, 349 35, 36, & 37. 0n_ a /'Cow J3asis'^« P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n -- 12 o r 80*0/£« D o u b t f u l " " " " — 2 o r 13.3#* N e g a t i v e " " " '* — 1 o r 6.7%. O j r ^ j a ^ u a r t e r B a s i s " . E i g h t e e n (±8) a p p a r e n t l y n o r m a l q u a r t e r s (no i n -j u r i e s and no p h y s i c a l l y a b n o r m a l m i l k p r o d u c e d a t any t i m e , i n c l u d i n g t i m e o f s a m p l i n g ) . P o s i t i v e C o n c l u s i o n s r e O s t i t i s I n f e c t i o n — 5i" o r 30.5%» D o u b t f u l " »• ~ " 11 — 5 o r 27*8%. N e g a t i v e " " " 11 ~ 7|- o r 41*7%. • 60. Group 3 - C o n t i n u e d T h i r t y - s e v e n (37) q u a r t e r s w i t h a m a s t i t i s h i s t o r y ( h a v i n g a t some t i m e p r o d u c e d p h y s i c a l l y a b n o r m a l m i l k ) , b u t •which a t t i m e o f s a m p l i n g were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k f r o m a l l - f o u r q u a r t e r s ) . P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n — 22 o r 59«5%» D o u b t f u l " " " " e>| o r 17.6%. N e g a t i v e H " " " — &§ o r 22. Grjpjxg. j 4 . Cows w i t h a m a s t i t i s h i s t o r y w h i c h a t t i m e o f samp-l i n g were s e c r e t i n g m i l k t h a t a p p e a r e d p h y s i c a l l y a b n o r m a l * T h e r e a r e n i n e (9) cows i n t h i s g r o u p : -N o s . 5, 6 , 1 1 , 12, 27, 2 8 , 4 1 , 49 and 50 . On a "Cow j B a s l s " . P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n 9 o r 100^» D o u b t f u l " " " " -- 0-N e g a t i v e " " " " 0 - - - - -On a, 11 Q.uarter Basia' 1,. T w e n t y - f i v e (25) a p p a r e n t l y n o r m a l q u a r t e r s (no i n -j u r i e s and no p h y s i c a l l y a b n o r m a l . m i l k p r o d u c e d a t - a n y t i m e s i n c l u d i n g t i m e o f s a m p l i n g ) * P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n -- yjr o r 38%. D o u b t f u l " " " " — 7f o r 30^.. N e g a t i v e " " " " 8 o r 3 2 f . 6 1 . Group 4 - C o n t i n u e d One i n j u r e d q u a r t e r w h i c h h a d n o t b e e n known t o p r o d u c e o t h e r t h a n a p p a r e n t l y n o r m a l m i l k . A P o s i t i v e C o n c l u s i o n r e M a s t i t i s I n f e c t i o n . T en (10) q u a r t e r s w h i c h a t t i m e o f s a m p l i n g w e re s e c r e t i n g m i l k t h a t a p p e a r e d p h y s i c a l l y a b n o r m a l . P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n -- 9iir o r 95%« D o u b t f u l " ' " " " — -I o r N e g a t i v e 11 " 11 " — 0 — — — P i n a l C l a s s i f i c a t i o n o f a l l Cows and a l l Q u a r t e r s . T e s t e d A c c o r d i n g t o P o s i t i v e , D o u b t f u l and N e g a t i v e C o n c l u s i o n s B e g a r d i n g M a s t i t i s I n f e c t i o n . . On a "Cow B a s i s " . F i f t y - t h r e e (53) cows were t e s t e d . P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n — 39X o r 74, D o u b t f u l " " <> » — 10 o r 18,9%. F e g a t i v e " " " " - 3 i o r 6 Onja_Jl£!mrter B a s i s " . Two h u n d r e d a n d s e v e n (207) q u a r t e r s were t e s t e d . ( S a m p l e s w ere n o t o b t a i n a b l e f r o m f i v e (5) q u a r t e r s o f t h e cows t e s t e d due t o a n a d v a n c e d s t a g e o f m a s t i t i s i n f e c t i o n . Cow No. 13 had o n l y one f u n c t i o n i n g q u a r t e r , RP» Cow No. 19 had o n l y two f u n c t i o n i n g q u a r t e r s . , KF and L P . ) P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n 76-J o r 36»9%. D o u b t f u l 11 " " " — 52f o r 25.4%. N e g a t i v e " " " " — 78 o r 37*7%. 62. I t i s s e e n t h a t a p p a r e n t l y n o r m a l q u a r t e r s were f o u n d I n e a c h o f t h e f o u r g r o u p s o f cows, A f i n a l c l a s s i f i -c a t i o n o f a l l a p p a r e n t l y n o r m a l q u a r t e r s i n Groups 1- 2, 3, 'and 4, a c c o r d i n g t o p o s i t i v e , d o u b t f u l and n e g a t i v e c o n c l u -s i o n s r e m a s t i t i s i n f e c t i o n i s h e r e p r e s e n t e d ; T o t a l Number o f s u c h Q u a r t e r s — 1 3 6 , P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n -- 35 o r 2^.7%, D o u b t f u l " " " " 40£ o r 2<)»8%. N e g a t i v e " " " 11 — 6Of o r 44.5^, Q u a r t e r s w h i c h had been i n j u r e d a t some t i m e b u t w h i c h were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k w e re f o u n d i n .Groups 2 and 4. A f i n a l c l a s s i f i c a t i o n o f t h e s e q u a r t e r s a c -c o r d i n g t o p o s i t i v e j d o u b t f u l a n d n e g a t i v e c o n c l u s i o n s r e mas-t i t i s i n f e c t i o n i s h e r e p r e s e n t e d °. T o t a l Number o f s u c h Q u a r t e r s -- 20 P o s i t i v e C o n c l u s i o n s r e M a s t i t i s I n f e c t i o n -- 10 o r 41.7%. D o u b t f u l " " " " — 5 o r 20.8jf. N e g a t i v e " " " " 9 o r 37. A l l q u a r t e r s w i t h a m a s t i t i s h i s t o r y ( h a v i n g a t some t i m e p r o d u c e d p h y s i c a l l y a b n o r m a l m i l k , b u t w h i c h a t t i m e o f s a m p l i n g were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k f r o m a l l f o u r q u a r t e r s ) a r e o f c o u r s e c o n t a i n e d i n t h e summary o f Group 3 > T a b l e No. 4. Any q u a r t e r s s e c r e t i n g a b n o r m a l m i l k a r e m e n t i o n e d i n t h e summary o f Group 4, T a b l e No. 4. 63. D i s c u s s i o n Of T a b l e l o . 4 B e f o r e d i s c u s s i n g t h e r e s u l t s p r e s e n t e d i n t h e sum-mary o f T a b l e No. 4, i t must o f c o u r s e be c l e a r l y u n d e r s t o o d t h a t a " p o s i t i v e " c o n c l u s i o n r e m a s t i t i s i n f e c t i o n f r o m any one q u a r t e r o f a cow, w h e t h e r o f Groups 1. 2, 3 o r 4, c l a s s e s t h a t cow a s h a v i n g a m a s t i t i s i n f e c t i o n . S i m i l a r l y , a " d o u b t f u l " c o n c l u s i o n f r o m any one q u a r -t e r o f a cow p l a c e s t h a t cow i n t h e d o u b t f u l c l a s s . I t I s t h u s s e e n t h a t t o be f i n a l l y t e r m e d a n o r m a l a n i m a l s a cow, o f n e c e s s i t y , must have " n e g a t i v e " c o n c l u s i o n s r e m a s t i t i s i n f e c t i o n f r o m a 11 f o u r q u a r t e r s . R e f e r r i n g now t o t h e summary o f T a b l e No. 4, i t i s s e e n t h a t o f t h e e i g h t cows w h i c h one w o u l d c o n s i d e r n o r m a l (Group l ) f b o t h f r o m c a s e h i s t o r i e s and f r o m t h e a p p e a r a n c e o f t h e m i l k a s drawn, 4-3.7% "were c l a s s e d a s h a v i n g a m a s t i t i s i n f e c t i o n . The p e r c e n t a g e o f cows g i v i n g a " p o s i t i v e " c o n c l u s i o n r e m a s t i t i s i n f e c t i o n i n c r e a s e s t o 71 • 4$ i n t h e ca.se o f t h e t w e n t y - o n e cows w h i c h had had i n j u r i e s t o q u a r t e r s (Group 2 ) s b u t w h i c h had a l w a y s p r o d u c e d m i l k t h a t a p p e a r e d n o r m a l f r o m b. p h y s i c a l e x a m i n a t i o n . The f i f t e e n cows w h i c h had a t some t i m e p r o d u c e d p h y s i c a l l y a b n o r m a l m i l k b u t w h i c h were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k a t t i m e o f s a m p l i n g (Group 3)» were f o u n d t o be 80/£ p o s i t i v e r e m a s t i t i s i n f e c t i o n . I n t h e f i n a l g r o u p o f n i n e cows (Group 4 ) , s e c r e t i n g p h y s i c a l l y a b n o r m a l m i l k , 100% were c o n s i d e r e d t o have a 64. m a s t i t i s , i n f e c t i o n . T h u s i n t h e f o u r g r o u p s t e s t e d , m a s t i t i s i n f e c t i o n i n c r e a s e d f r o m 4 3 , 7 % t o 7 1 , 4 % t o 71*4% t o 8o«0% t o 100%. W i t h r e g a r d t o cows p l a c e d i n t h e " d o u b t f u l " c l a s s , i t i s s e e n t h a t no r e a l l y d e f i n i t e c o n c l u s i o n s can be drawn, e x c e p t i n so f a r a s i n d i c a t i n g t h a t t h e s e cows a r e open t o t h e s u s p i c i o n o r a s s u m p t i o n t h a t a m a s t i t i s c o n d i t i o n may d e v e l o p and r e a c h a d e f i n i t e l y d e t e c t a b l e stage« The p e r c e n t a g e o f " d o u b t f u l " cows d e c r e a s e d f r o m G r o u p s 1 t o 4 , a s t h e p e r c e n t a g e , o f . m a s t i t i s i n f e c t e d cows i n c r e a s e d , i n d i c a t i n g t h a t a s a b n o r m a l i t i e s i n c a s e h i s t o r i e s bega.n t o a p p e a r p r o c e e d i n g f r o m Group i t o Group 4 S an i n c r e a -s i n g p e r c e n t a g e o f cows were p l a c e d i n t h e c l a s s t e r m e d " p o s -i t i v e " r e m a s t i t i s i n f e c t i o n . I t i s r e a s o n a b l e t o assume t h e r e f o r e , t h a t t h o s e cows p l a c e d i n t h e " d o u b t f u l " c l a s s may have a m i l d e r f o r m o f m a s t i t i s i n f e c t i o n . I t i s o f i n t e r e s t t o n o t e t h a t of t h e e i g h t cows c o n s i d e r e d t o be n o r m a l f r o m t h e v i e w p o i n t . o f " c a s e h i s t o r y " a nd p h y s i c a l c o n d i t i o n o f m i l k a t t i m e of s a m p l i n g (Group 1 ) , o n l y 1 8 . 8 % gave " n e g a t i v e " c o n c l u s i o n s r e m a s t i t i s i n f e c t i o n . T h i s p e r c e n t a g e i s d e c r e a s e d t o 4 . 8 % i n t h e c a s e o f cows w h i c h had had i n j u r i e s t o q u a r t e r s a t some t i m e b u t w h i c h had a l w a y s p r o d u c e d a p p a r e n t l y n o r m a l m i l k , (Group 2 ) , and. t o 6J]% i n t h o s e cows w h i c h had p r o d u c e d a b n o r m a l m i l k a t some t i m e b u t w h i c h were s e c r e t i n g a p p a r e n t l y n o r m a l m i l k a t t h e 65. t i m e o f s a m p l i n g (Group 3)• Somewhat d i f f e r e n t p e r c e n t a g e s a r e a r r i v e d a t i n c l a s s i f y i n g t h e q u a r t e r s t e s t e d due t o t h e f a c t t h a t cows c o n -. s i d e r e d a s " p o s i t i v e " r e m a s t i t i s i n f e c t i o n may have one o r more q u a r t e r s t h a t a r e c l a s s e d as ' d o u b t f u l ' o r "negative's, Sim-i l a r l y , cows i n t h e " d o u b t f u l " c l a s s may have q u a r t e r s c l a s -s e d a s " n e g a t i v e 1 ! Of a l l a p p a r e n t l y n o r m a l q u a r t e r s t e s t e d i n t h e f o u r g r o u p s , 25.7?^ were c o n s i d e r e d t o h a v e a m a s t i t i s i n f e c t i o n . I n t h e i n j u r e d q u a r t e r s (Group 2, and one- i n Group 4) , 41 „ 7^ o f t h o s e t e s t e d w e re c o n s i d e r e d t o have a " p o s i t i v e " c o n c l u -s i o n r e m a s t i t i s i n f e c t i o n . Q u a r t e r s w h i c h had had a m a s t i t i s h i s t o r y ( a s s e e n i n Group 3), were 59«5/£ p o s i t i v e r e m a s t i t i s i n f e c t i o n , and t h o s e q u a r t e r s s e c r e t i n g a b n o r m a l m i l k (Group 4 ) , were f o u n d t o have a 95% m a s t i t i s i n f e c t ion.. W i t h r e g a r d t o " d o u b t f u l " c o n c l u s i o n s r e m a s t i t i s i n f e c t i o n i n q u a r t e r s i n t h e f o u r g r o u p s , t h e p e r c e n t a g e o f t h e s e d e c r e a s e d a s t h e p e r c e n t a g e o f p o s i t i v e c o n c l u s i o n s i n -c r e a s e d . Of t h e a p p a r e n t l y n o r m a l q u a r t e r s , 29.8% were c o n -s i d e r e d d o u b t f u l ? o f t h e i n j u r e d q u a r t e r s , 20.8%; o f t h e mas-t i t i s h i s t o r y q u a . r t e r s , 17 »6/£j and o f t h e q u a r t e r s p r o d u c i n g a b n o r m a l m i l k , 5/^ * The p e r c e n t a g e o f " n e g a t i v e " c o n c l u s i o n s i n e a c h g r o u p on a q u a r t e r b a s i s , , d e c r e a s e d a l s o a s t h e " p o s i t i v e s " i n c r e a s e d . Of t h e a p p a r e n t l y n o r m a l q u a r t e r s , 44 .5'? were c o n -66. s i d e r e d f r e e o f m a s t i t i s i n f e c t i o n , I n j u r i e s t o q u a r t e r s r e -duced t h e number f o u n d " n e g a t i v e " t o 37.5%, Of t h e q u a r t e r s w i t h a. m a s t i t i s h i s t o r y s 22 ,<)% were c o n s i d e r e d " n e g a t i v e " - w h i l e none o f t h e q u a r t e r s • s e c r e t i n g a b n o r m a l m i l k a t t i m e o f s a m p l i n g came i n t h e " n e g a t i v e " c l a s s . ' ^SIli.JLL G-gi n c-g...- a nd - A p p l i c a t i o n o f M a t e r i a l I n T a b l e ITo, 4 & A number of p r o b l e m s a r i s e f r o m t h e f o r e g o i n g d i s -c u s s i o n * One o f t h e r a t h e r s u r p r i s i n g r e s u l t s i n t h i s i n v e s -t i g a t i o n i s t h e h i g h i n c i d e n c e (43,7%\ o f m a s t i t i s f o u n d i n cows b e l i e v e d t o be a b s o l u t e l y n o r m a l . O n l y 18.8 °/Q were r e -g a r d e d a s e n t i r e l y f r e e o f i n f e c t i o n -- t h e b a l a n c e b e i n g i n t h e d o u b t f u l c l a s s . I t needs t o be remembered t h a t t h e r e were o n l y e i g h t (8) cows i n t h i s g r o u p and t h a t a l a r g e r number m i g h t g i v e a d i f f e r e n t , and c e r t a i n l y a t r u e r c o n c e p t i o n o f t h e i n c i d e n c e o f m a s t i t i s i n cows b e l i e v e d t o be f r e e o f . t h e d i s e a s e . An a n a l y s i s o f T a b l e No, 4 d i s c l o s e s a c o n d i t i o n w h i c h I s v e r y d e f i n i t e l y r e l a t e d t o h e r d management, b a r n s a n i t a t i o n and m a s t i t i s p r e v e n t i o n a nd c o n t r o l . I t I s seen that, o f t h e t w e n t y - o n e (21) cows w i t h i n j u r i e s t o u d d e r o r t e a t s * b u t w h i c h have n o t been known t o p r o d u c e o t h e r t h a n a p p a r e n t l y n o r m a l m i l k , 7^-o^% were c o n s i d e r e d a s h a v i n g a. m a s t i t i s i n f e c -t i o n , a l m o s t a q u a r t e r w e re c l a s s e d a s " d o u b t f u l " and l e s s t h a n 5% were " n e g a t i v e " . One ( l ) cow, o n l y , p r o v e d t o be e n t i r e l y f r e e o f m a s t i t i s i n f e c t i o n . 67. Of t h e f i f t e e n (15) cows w i t h a m a s t i t i s h i s t o r y b u t a p p a r e n t l y r e c o v e r e d , a s l i g h t l y h i g h e r p e r c e n t a g e {Qo.0%) were c o n s i d e r e d " p o s i t i v e " , f e w e r cows were i n t h e " d o u b t f u l " ' c l a s s and a g a i n o n l y one cow out o f t h e f i f t e e n (15) b e l i e v e d t o be s e c r e t i n g norma.! m i l k . • wa,s a c t u a l l y p e r f o r m i n g t h i s f u n c t i o n . The r e l a t i o n s h i p o f t h e s e f i n d i n g s t o h e r d manage-ment i s r e a d i l y a p p a r e n t . The d a i r y m a n who r e g a r d s s u c h cows as e i t h e r f r e e o f m a s t i t i s i n f e c t i o n o r a s h a v i n g r e g a i n e d a p e r m a n e n t o r even t e m p o r a r y n o r m a l i t y (and t h e authors„ as p r e v i o u s l y s t a t e d , b e l i e v e most d a i r y m e n a r e o f t h i s class)„ t r e a t s them a s n o r m a l cows. S u c h a p r o c e d u r e c a n n o t h e l p b u t . c o n t r i b u t e t o a s t i l l w i d e r s p r e a d o f t h i s c o n t a g i o u s d i s e a s e . I n v i e w o f t h e f a c t t h a t f e w h e r d s ( i n d e e d , i f a n y ) . no m a t t e r how c a r e f u l l y managed, can be k e p t f r e e o f u d d e r o r t e a t i n -j u r i e s j . t h e a u t h o r s s u g g e s t t h a t t h e use o f an e f f e c t i v e d i s -i n f e c t i n g s o l u t i o n f o r t h e cows u d d e r s and f o r t h e m i l k e r s ' hands ( o r t h e m i l k i n g machine) m i g h t w e l l be c o n s i d e r e d an e s -s e n t i a l s a n i t a r y p r e c a u t i o n as e a c h cow i s m i l k e d . The h i g h i n c i d e n c e o f m a s t i t i s f o u n d i n cows w h i c h had t e a t o r u d d e r i n j u r i e s b u t no known p r o d u c t i o n of a b n o r m a l m i l k , may be e x p l a i n e d b y a r e c e n t r e p o r t f r o m t h e New Y o r k S t a t e A g r i c u l t u r a l E x p e r i m e n t S t a t i o n by H u c k e r ('7)« He s t a t e s i n p a r t : "From t h e f o r e g o i n g r e s u l t s i t i s o b v i o u s t h a t cows w h i c h have p a s s e d t h r o u g h one o r more l a c t a t i o n p e r i o d s n e a r l y a l w a y s , i f n o t a l w a y s , h a r b o r some member o f t h e m a s t i t i s 68. s t r e p t o c o c c u s g r o u p i n t h e u d d e r t i s s u e . Whether t h e p r e s -. ence o f t h e m a s t i t i s s t r e p t o c o c c i i n t h e s e u d d e r s w h i c h have p a s s e d t h r o u g h a l a c t a t i o n p e r i o d c o n s t i t u t e s an i n f e c t i o n i s , a m a t t e r o f d e f i n i t i o n o f t e r m s . C e r t a i n l y no e v i d e n c e o f p a t h o l o g i c a l change i n t h e t i s s u e s o r o f a n i n f e c t i o n was a p p a r e n t . The o r g a n i s m s a r e a p p a r e n t l y p r e s e n t i n a dormant s t a t e a n d a r e c o m p l e t e l y s u p p r e s s e d ; a s f a r a s any a c t i v i t y i s c o n c e r n e d , by t h e r e s i s t a n c e o f t h e u d d e r t i s s u e . " H u c k e r s t a t e s t h a t t h e o r g a n i z m s p r o b a b l y i n v a d e t h e u d d e r t i s s u e t h r o u g h t h e t e a t c a n a l and t h a t t h e n o r m a l hab-i t a t o f m a s t i t i s s t r e p t o c o c c i needs i n t e n s i v e s t u d y . He s a y s , f u r t h e r , t h a t d a i r y m e n o b s e r v e t h a t a n u d d e r b r u i s e o r i n -j u r y i s f r e q u e n t l y f o l l o w e d by an a c u t e c a s e o f m a s t i t i s i n a p r e v i o u s l y n o r m a l cow* These o b s e r v a t i o n s a p p e a r t o be i n harmony w i t h t h e r e s u l t s o b t a i n e d by t h e a u t h o r s on t h i s i n v e c t i g a t i o n . C e r -t a i n l y i n h e r d s where some m a s t i t i s i s known t o be p r e s e n t one w o u l d c o n c l u d e f r o m t h e a b o v e t h a t a l l , o r p r a c t i c a l l y a l l s cows have t h e o r g a n i s m s p r e s e n t i n t h e udder, t i s s u e , ( H u c k e r (7) e v e n f o u n d s t r e p t o c o c c i m a s t i t i s organisms, p r e s e n t i n 47% o f t h e u d d e r s o f v i r g i n h e i f e r s ) „ A t e a t i n j u r y , c h i l l o r b r u i s e may weaken t h e n o r m a l r e s i s t a n c e o f the u d d e r t i s s u e and. t h e o r g a n i s m s commence t o s e t up an i n f e c t i o n , , T h i s may be a c u t e i n n a t u r e , o r i t may t a k e c o n s i d e r a b l e t i m e t o d e v e l o p . E v e n t u a l l y , h owever, a r a t h e r s e r i o u s c o n d i t i o n u s u a l l y r e s u l t s , w i t h m i l k p r o d u c t i o n r e d u c e d i n b o t h q u a n t i t y 69. a n d q u a l i t y . The i m p o r t a n c e o f so managing a d a i r y h e r d t h a t u d d e r and t e a t i n j u r i e s a r e r e d u c e d t o a minimum, and t h a t " m a s t i t i s -. s p r e a d e r s " a r e d e t e c t e d and. e f f e c t i v e l y p r e v e n t e d f r o m i n f e c -t i n g h e a l t h y cows, i s r e a d i l y a p p a r e n t . The a u t h o r s w i s h t o r e f e r " b r i e f l y t o t h e p r o b l e m o f t h e d e t e c t i o n o f v a r i o u s s t a g e s o f m a s t i t i s i n f e c t i o n s . From t h e f o r e g o i n g d i s c u s s i o n i t w o u l d seem t o be d e f i n i t e l y e s t a b -l i s h e d t h a t much o f t h e a p p a r e n t l y n o r m a l m i l k s e c r e t e d by d a i -r y cows m i g h t be more c o r r e c t l y t e r m e d " m a s t i t i s m i l k " , H u c k e r (7) e s t i m a t e s t h a t "a s u r v e y o f any group o f cows w o u l d p r o b a b l y show t h a t 30 t o 5° p e r c e n t o f t h e a n i m a l s d i s c h a r g e s t r e p t o c o c c i i n t h e i r m i l k " . I t a p p e a r s t h a t c o n d i t i o n s e x i s t i n t h e b o v i n e u d d e r r a n g i n g f r o m cows w i t h "dormant" s t r e p t o -c o c c i p r e s e n t i n t h e u d d e r t i s s u e t o t h o s e a n i m a l s i n w h i c h a n i n f e c t i o n has d e v e l o p e d t o t h e e x t e n t o f c o m p l e t e l y c h a n g i n g t h e p h y s i c a l , b a c t e r i o l o g i c a l and c h e m i c a l n a t u r e o f t h e m i l k , o r o f so c h a n g i n g t h e c o n d i t i o n o f t h e udder t h a t i t may be-come n o n - f u n c t i o n i n g o r " b l i n d " i n one o r more q u a r t e r s , . I t seems r a t h e r d i f f i c u l t , t o s a y j u s t when a n i n f e c -t i o n i s p r e s e n t . The more s e n s i t i v e t h e t e s t and t h e g r e a t e r t h e number o f t e s t s a p p l i e d , t h e e a r l i e r w o u l d an i n f e c t i o n be i n d i c a t e d . P o s s i b l y any d e p a r t u r e f r o m t h e n o r m a l by a w e l l -e s t a b l i s h e d t e s t i n d i c a t e s t h e b e g i n n i n g o f an i n f e c t i o n . To be on t h e " s a f e " s i d e , t h e a u t h o r s i n t h i s i n v e s t i g a t i o n have r e p r e s e n t e d 25.4%' o f t h e q u a r t e r s (l8«9% o f t h e cows) t e s t e d 70. a s " d o u b t f u l " r e m a s t i t i s i n f e c t i o n . I t may be t h a t t h i s w h o l e " d o u b t f u l g r o u p " i s c h a r a c t e r i z e d b y a v e r y m i l d f o r m o f i n f e c t i o n w h i c h may d e v e l o p , i n t i m e , i n t o a more d e f i n -. I t e l y d e t e c t a b l e c o n d i t i o n . T h i s p r o b l e m I s b r i e f l y o u t l i n e d b e c a u s e a r e v i e w o f t h e l i t e r a t u r e i n d i c a t e s a f a i r l y w i d e d i v e r g e n c e o f o p i n -i o n amongst l e a d i n g i n v e s t i g a t o r s r e g a r d i n g t h e r e l i a b i l i t ; / , s e n s i t i v e n e s s and v a l u e o f t h e d i f f e r e n t t e s t s , and b e c a u s e ' what c o n s t i t u t e s an i n f e c t i o n i s , as. H u c k e r ( 7 ) s a y s , " „. , . a m a t t e r o f d e f i n i t i o n o f t e r m s 1 1 . I t w o u l d a p p e a r t h a t f u r t h e r r e s e a r c h i n t h i s r e g a r d i s e s s e n t i a l i n o r d e r t o e s t a b l i s h a more d e f i n i t e f o u n d a t i o n upon w h i c h t o b a s e a programme o f e f f e c t i v e m a s t i t i s c o n t r o l o r e l i m i n a t i o n . U n t i l s u c h t i m e a s t h i s i s a c c o m p l i s h e d , a p r o c e d u r e o f t e s t i n g and i n t e r p r e -t a t i o n somewhat s i m i l a r t o t h a t used f o r t h e cows g r o u p e d i n T a b l e F o . 3 w o u l d , i n t h e a u t h o r s * o p i n i o n , g i v e a r e a s o n a b l y a c c u r a t e p i c t u r e o f m a s t i t i s c o n d i t i o n s and c o n s t i t u t e a f a i r l y s o und b a s i s f o r t h e p r a c t i c a 1 and e c o n o m i c a l c o n t r o l o f t h e d i s e a s e * A q u e s t i o n n a t u r a l l y a r i s e s a s t o t h e s u i t a b i l i t y o f cow's m i l k f o r human c o n s u m p t i o n . T h e r e i s l i t t l e o r no e v i d e n c e t o i n d i c a t e t h a t t h e m i l d e s t f o r m s o f m a s t i t i s I n d a i r y cows r e s u l t i n t h e p r o d u c t i o n o f m i l k t h a t i s u n s u i t e d . f o r t h e f l u i d m a r k e t i n e i t h e r t h e raw o r p a s t e u r i z e d state«-D a h l b e r g a n d a s s o c i a t e s (8) f o u n d no s i g n i f i c a n t changes i n t h e c h e m i c a l c o m p o s i t i o n o f m i l k drawn f r o m cows w i t h l a t e n t 71 I n f e c t i o n s o f m a s t i t i s so l o n g as t h e m i l k r e m a i n e d n o r m a l i n i t s p h y s i c a l a p p e a r a n c e and c o n c l u d e d t h a t i f a l l a b n o r m a l m i l k f r o m i n f l a m e d , c o n g e s t e d , o r i n j u r e d q u a r t e r s i s e x c l u -d e d , m i x e d m i l k w i l l be e s s e n t i a l l y n o r m a l i n c h e m i c a l com-p o s i t i o n , The b a c t e r i o l o g i c a l r e l a t i o n s h i p t o p u b l i c h e a l t h o f a p p a r e n t l y n o r m a l m i l k 5 however, i s . more d e b a t a b l e . Out-b r e a k s o f s e p t i c s o r e t h r o a t have been t r a c e d t o m i l k s u p p l i e s I n a r e a s where s u c h o u t b r e a k s have been of a s e r i o u s n a t u r e , c o m p u l s o r y p a s t e u r i z a t i o n has u s u a l l y been e n f o r c e d . C l a r k (9) f r o m a s t u d y o f t h e p r o d u c t i o n o f u n d e s i r a b l e o d o r s i n raw m i l k by s p e c i f i c o r g a n i s m s a s s o c i a t e d , w i t h u d d e r i n f e c -t i o n s , c o n c l u d e d , t h a t m a s t i t i s i n d a i r y c a t t l e : may be a s s o c -i a t e d w i t h t o x i n - p r o d u c i n g s t a p h y l o c o c c i , c a p a b l e o f c a u s i n g g a s t o e n t e r i t i s i n man. and t h a t o r g a n i s m s c a p a b l e o f p r o d u c i n g u n d e s i r a b l e o d o r s i n m i l k may l o c a l i z e i n m a s t i t i s - i n f e c t e d u d d e r s . G-wa,tkin, and a s s o c i a t e s ( 1 0 ) found, t h a t i n one hundred and f o r t y - t h r e e c a s e s o f m a s t i t i s , t h i r t y were t h e r e s u l t o f s t a p h y l o c o c c i c i n f e c t i o n , , T o x i n was p r o d u c e d hy 67*A% o f t h e s t r a i n s r e c o v e r e d , f r o m s t a p h y l o c o c c i c m a s t i t i s and from. 2 0 * 0 % o f t h e s t r a i n s i s o l a t e d f r o m n o r m a l u d d e r s . D r , C. D. M c G i l -v a r y (11) i n d i s c u s s i n g m a s t i t i s i n cows c o n c l u d e s t h a t 11 ... i n o r d e r t o p r o t e c t p u b l i c h e a l t h t h e m i l k f r o m m a s t i t i s a f f e c -ted, h e r d s s h o u l d n o t be u s e d f o r human c o n s u m p t i o n u n l e s s i t i s p r o p e r l y p a s t e u r i z e d " . The a u t h o r s a r e o f t h e o p i n i o n t h a t u n l e s s e f f e c t i v e s t e p s c a n be t a k e n t o e n s u r e t h a t h e r d s s u p p l y i n g raw m i l k 72. f o r f l u i d c o n s u m p t i o n a r e m a i n t a i n e d i n a r e l a t i v e l y m a s t i t i s -f r e e c o n d i t i o n , , c o m p u l s o r y p a s t e u r i z a t i o n i s n e c e s s a r y i n o r d e r t o s a f e g u a r d p u b l i c h e a l t h . ' ' CHLORIM; CONTENT, O F MILK SAMPLES AS RSIAT2D TO HOTIS US ST T a b l e No. 5* a s n a s been mentioned., r e p r e s e n t s a s t u d y , i n t a b l e f o r m , o f t h e r e s u l t s o f t h e c h l o r i n e and H o t i s t e s t r e s u l t s a s r e v e a l e d i n T a b l e s No. 2 and No. 3« I h e mat-e r i a l i s p r e s e n t e d i n t h r e e ' w a y s ; I n P a r t 1, " C h l o r i n e Read-i n g s " a r e c l a s s i f i e d . I n t o " H o t i s G r o u p s " ; i n P a r t 2, " H o t i s R e a d i n g s " a r e c l a s s i f i e d i n t o " C h l o r i n e C o n t e n t . G r o u p s " j and I n P a r t 3 ? 1 ' ^ o t i s T e s t r e s u l t s ' 1 a r e c o r r e l a t e d t o e a c h o f f o u r d i f f e r e n t s t a n d a r d s f o r n o r m a l c h l o r i n e c o n t e n t * "HIa.ch p a r t I s s u b d i v i d e d i n t o t h r e e s e c t i o n s ; S e c t i o n A, b a s e d on m a t e r i a l i n T a b l e No, 2.; S e c t i o n B, b a s e d on m a t e r i a l i n T a b l e No. 3? an d S e c t i o n C, b a s e d on co m b i n e d m a t e r i a l i n T a b l e s No. 2 and No. 3. A. s h o r t d i s c u s s i o n f o l l o w s t h e p r e s e n t a t i o n o f the m a t e r i a l i n t a b l e f o r m . 73. 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O V+ VO E>-t>- VO ON CO O t c M n , ' tr\ tr\ flVO O N Hr l r \ O H ; -o ro** . c » H fl cM ro<* r o o o c\l r o e & s H O O r H r o O ro^i- O- VO CO CO rH r O § fl ro-* IN- .rH CO fl CO CO CQ CQ . fl co CD CD i> ZS f> l> -ri <t-H CD CD CO CD -rH • H - H P -P • > j> {> f> H J , 1 P P ••~! fl C - H - H -rH >H crj fjd .H - H CQ 3 CO P P +-> + 3 M) S3; ra CQ o g P crj « crj crj crj CD J * H 0 o PH H O tiO^S bjj co tjO t i o f l flfl. fH CD r«-f CD crj CD CD * - i fl fl fl <; fl flflfl CD - T fl H 1 1 1 ( 1—; fl fl PH < < fl fl fl fl <j; 84.. D i s c u s s i o n of T a b l e Ho, 5 P a r t 1 -~ C l a s s i f i c a t i o n o f " C h l o r i n e R e a d i n g s " i n t o " H o t i s G r o u p s " I t i s r e a d i l y seen f r o m a s t u d y o f t h e " c h l o r i n e r e a d i n g s " p r e s e n t e d i n t h e v a r i o u s " H o t i s G r o u p s " i n S e c t i o n s A and B o f P a r t . 1, t h a t a w i d e range o f r e a d i n g s o c c u r s . ' T h i s i s t h e main p u r p o s e s e r v e d by t h e s e two s e c t i o n s . S e c t i o n C 5 w h i c h i s a co m b i n e d summary o f S e c t i o n s A and B,. r e q u i r e s c a r e f u l s t u d y . R e f e r r i n g t o t h e " A v e r a g e C h l o r i n e " c o l u m n i t i s a p p a r e n t t h a t f o r each H o t i s ' r e a d i n g , t h e a v e r a g e c h l o r i n e c o n t e n t i s c o n s i s t e n t l y l o w e r f o r t h e m i l k s a m p l e s o f cows i n T a b l e 2 a s compared t o T a b l e 3» T h i s may be due t o a d i f f e r e n t degree o f m a . s t i t i s i n f e c t i o n i n t h e two groups, o f cows o r i t may be a c c o u n t e d f o r by t h e d i f f e r -ence i n numbers o f cows ( t h i r t e e n i n T a b l e Ho, 2, and t w e n t y -s e v e n i n T a b l e No »•3)• The a v e r a g e c h l o r i n e c o n t e n t f o r a l l " H o t i s P o s i t i v e s " i s 0.1542#. F o r a l l " H o t i s D o u b t f u l s " i t i s 0.146'1$ and f o r " F o r m a l (H) n e g a t i v e s " , .0.1461^. F o r " A l l Negatives''' t h e c o n t e n t i s 0.1511^ due t o t h e h i g h c o n t e n t o f 0.1755% i n t h e " A l l o t h e r n e g a t i v e s " g r o u p . The h i g h c o n t e n t o f c h l o r i n e i n t h i s l a t t e r g r o up may be e x p l a i n e d by an udder i n f e c t i o n due t o a. c a u s a l o r g a n i s m o t h e r t h a n s t r e p t o c o c c u s m a s t i t i d i s (and hence n o t g i v i n g a P o s i t i v e H o t i s r e a c t i o n ) o r i t may be t h a t t h e i n f e c t i o n was v e r y s e v e r e and t h e c h a r a c t e r of" t h e m i l k so a l t e r e d t h a t a P o s i t i v e H o t i s r e a c t i o n was n o t discernable» I n t h e e x p e r i e n c e o f t h e a u t h o r s w i t h t h e H o t i s t e s t a t y p i c a l l y p o s i t i v e r e a c t i o n i s r a r e l y obtained, w i t h a 85. m i l k sample t h a t i s v e r y c l o t t y and serous« I n t h e "maximum" and. "minimum" columns t h e c h l o r i n e p e r c e n t a g e s a r e so d i s t r i b u t e d a s t o c h a l l e n g e e x p l a n a t i o n , e x c e p t on t h e b a s i s m e n t i o n e d above i n d i s c u s s i n g t h e " a v e r -age" group,. P a r t . 2 — C l a s s i f i c a t i o n o f " H o t i s Rea.dings" i n t o " C h l o r i n e C o n t e n t G r o u p s " . a n d P a r t 3 C o r r e l a t i o n o f H o t i s t e s t r e s u l t s t o each o f f o u r d i f f e r e n t s t a n d a r d s f o r n o r m a l c h l o r i n e c o n t e n t . These two p a r t s o f T a b l e Ho* 5 i n d i c a t e t h e n e c e s -s i t y (and t h e d i f f i c u l t y ) insofar a s t h e H o t i s T e s t i s c o n c e r -n e d , o f e s t a b l i s h i n g a p r o p e r a r b i t r a r y c h l o r i n e c o n t e n t 9 above and b e l o w whi c h , m i l k may be c o n s i d e r e d as h a v i n g , o r b e i n g f r e e o f ? a. mastitis i n f e c t i o n . I n s t u d y i n g t h i s m a t e r i a l , howevers i t must be remembered, t h a t t h e H o t i s T e s t i s . d e s i g n e d t o de-t e c t t h e p r e s e n c e o f s t r e p t o c o c c u s m a s t i t i s , w hereas t h e c h l o r i n e t e s t i s r e g a r d e d a s a p p l i c a b l e t o a l l m a s t i t i s c o n d i -t i o n s , A f e w examples w i l l i l l u s t r a t e t h e i n t e r p r e t a t i o n w h i c h may be p l a c e d on t h e s e two p a r t s o f T a b l e Ho „ 5" In S e c t i o n A o f P a r t 2, i t i s se e n t h a t f a i r l y l a r g e p e r c e n t a g e s of H o t i s " A l l P o s i t i v e s " and " A l l n e g a t i v e s " f a l l I n e a c h o f t h e t h r e e i n t e r m e d i a t e " C h l o r i n e C o n t e n t G r o u p s " , I n S e c t i o n B 5 t h e same i s t r u e o f " A l l N e g a t i v e s " b u t t h e " A l l P o s i t i v e s " a r e largely c o n f i n e d t o t h e h i g h e r " C h l o r i n e G r o u p s " , I n S e c t i o n C 5 ( S e c t i o n s A and B combined) t h e r e s u l t more 86. n e a r l y p a r a l l e l s t h a t i n S e c t i o n B due t o t h e l a r g e r number o f cows r e p r e s e n t e d i n t h e l a t t e r s e c t i o n . P a r t 3 i n d i c a t e s t h e d e g r e e o f c o r r e l a t i o n b e t ween * t h e two t e s t s v e r y c l e a r l y . I t w i l l be n o t e d i n a l l t h r e e s e c -t i o n s t h a t i n o r d e r t o o b t a i n a r e a s o n a b l y s a t i s f a c t o r y c o r -r e l a t i o n i n t h e p o s i t i v e H o t i s r e a d i n g s i t i s n e c e s s a r y t o t h e l o w e r c h l o r i n e s t a n d a r d s . The d i f f i c u l t y i s , h owever, t h a t when t h e l o w e r s t a n d a r d s a r e t a k e n , a h i g h p e r c e n t a g e of t h e n e g a t i v e H o t i s r e a d i n g s a r e f o u n d w i t h i n t h e range o f what w o u l d t h e n be p o s i t i v e c h l o r i n e percentages« On t h e b a s i s o f t h e p o s i t i v e H o t i s r e a d i n g s , i t w o u l d a p p e a r t h a t a n o r m a l s t a n d a r d f o r c h l o r i n e i n t h e n e i g h b o r h o o d o f 0=1384% i s n e c e s -s a r y t o c o r r e l a t e s a t i s f a c t o r i l y w i t h H o t i s t e s t r e s u l t s . Us-i n g t h e n e g a t i v e H o t i s r e a d i n g s as a guide., a n o r m a l s t a n d a r d f o r c h l o r i n e i n t h e n e i g h b o r h o o d o f 0 , 1 5 % seems t o be d e s i r a b l e I t i s p o s s i b l e , o f c o u r s e , t h a t n e g a t i v e H o t i s r e -s u l t s were o b t a i n e d f r o m a number o f m i l k s a m p l e s . g i v i n g a r e l a t i v e l y h i g h c h l o r i n e c o n t e n t , due t o a m a s t i t i s i n f e c t i o n c a u s e d by s t a p h y l o c o c c i , . Case ( 2 ) s i t w i l l be remembered 5 f o u n d t h a t s t a p h y l o c o c c i c a u s e a l o w - g r a d e i n f e c t i o n i n 39-0% o f t h e c a s e s o f m a s t i t i s , P a h l b e r g and a s s o c i a t e s (8) c o n -c l u d e d s however, t h a t i n l a t e n t i n f e c t i o n s o f mastitis» t h e c h l o r i n e c o n t e n t i s o n l y s l i g h t l y i n c r e a s e d and t h a t d i f f e r -e n c e s i n c h e m i c a l c o m p o s i t i o n o f m i l k t h a t r e m a i n e d n o r m a l i s no g r e a t e r t h a n v a r i a t i o n s i n c h e m i c a l c o m p o s i t i o n o f m i l k b e t ween two h e r d s o f t h e same b r e e d * T h i s c o n c l u s i o n w o u l d 87. seem t o i n d i c a t e t h a t t h e c h l o r i n e t e s t i s o f l i t t l e use i n d e t e c t i n g a m a s t i t i s ' c o n d i t i o n i n cows s e c r e t i n g a p p a r e n t l y n o r m a l m i l k . Shaw and a s s o c i a t e s (12) i n d i s c u s s i n g t h e r e l i a b i l -i t y o f s e l e c t e d t e s t s ( i n c l u d i n g c h l o r i n e ) f o r t h e d e t e c t i o n of m a s t i t i s , s t a t e ? "The t e s t s u s i n g t h e p e r c e n t a g e o f c h l o r - . i d e s and h y d r o g e n - i o n c o n c e n t r a t i o n were e x t r e m e l y u n r e l i a b l e i n d e t e c t i n g cows s u f f e r i n g f r o m c h r o n i c m a s t i t i s " . B l o o d and Ro?;lands (13) who ma.de a s p e c i a l s t u d y o f t h e c h l o r i n e c o n t e n t o f m i l k as an i n d i c a t i o n o f m a s t i t i s con-c l u d e d t h a t " . . . . t h e d e t e r m i n a t i o n o f t h e c h l o r i n e c o n t e n t o f m i l k i s n o t a r e l i a b l e method f o r t h e a s c e r t a i n m e n t o f t h e p r e s e n c e o r a b s e n c e o f m a s t i t i s i n d a i r y cows". On t h e o t h e r h a n d , Rose11, t h e c h i e f e x ponent o f t h e c h l o r i n e t e s t 5 i s o f h i g h s t a n d i n g i n h i s p r o f e s s i o n and h i s c l a i m s f o r t h e t e s t a r e s u p p o r t e d by a number o f o t h e r w o r k e r s . The a u t h o r s f e e l t h a t i n t h e i n v e s t i g a t i o n h e r e r e -c o r d e d , t h e r e i s n o t s u f f i c i e n t e x p e r i m e n t a l e v i d e n c e t o sup-p o r t d e f i n i t e c o n c l u s i o n s * The above d i s c u s s i o n o f t h e c h l o r -i n e t e s t , however, p r o v i d e s a n e x c e l l e n t example o f t h e l a c k o f u n i f i e d o p i n i o n r e g a r d i n g t h e m e r i t s o f t h e v a r i o u s t e s t s f o r d e t e c t i n g m a s t i t i s i n d a i r y c a t t l e * 88. SUMMARY A I D CONCLUSIONS: F i f t y - three (53) cov/s were tested i n each quarter of the udder to determine the presence or absence of a mastitis infection. Of these 74.5% were considered as having mastitis, 18.9% were given a doubtful interpret-ation, and 6,6% were considered free of the disease. Of the eight (8) cows regarded, previous to testing, as absolutely normal i n every respect, 45.7% were regarded as d e f i n i t e l y infected with mastitis end only one ( l ) of the eight (8) was e n t i r e l y free of infection. In the "udder or teat injury" group with no known production of abnormal milk at any time 71.4% of the twenty - one (21) were positive, 23.8% were doubtful, and again, only one was negative. The f i f t e e n (lj?) cows secreting apparently normal milk at time of sampling but which were known to have produced physically abnormal milk at some time were con-sidered to be 80.0% positive, 13.3% doubtful and 6.7% negative re mastitis infection. Of the nine (9) cows secreting physically abnormal milk at time of sampling a l l (100%) were found to be positive. This investigation indicates that mastitis i s an exceedingly prevalent disease In the dairy herds of B r i t i s h Columbia. The authors are of the opinion that i t i s of much commoner occurrence than i s generally 89. ST3MMABY AID CONCLUSIONS: ~ (continued.) realized. It i s f e l t that few, i f any, dairymen have a true conception of the mastitis condition of their own cattle and that many eows are regarded as normal which act wily, are affected by an early development of the disease. The authors conclude that the common procedure of semi-isolation of cows known to Toe producing physically abnormal milk and the milking of such cows l a s t , does not constitute satisfactory control. It i s suggested that more effective precautions he taken to control mastitis "because of the high incidence of the disease i i i cows secreting apparently normal milk. In the more advanced stages of infection, the tests employed correlated to a greater degree than i n the milder-cases. The correlation between the Hotis and Chlorine tests, while excellent i n some instances, was negative i n others and on the average could be considered only.fair. The Dislac test i s considered of particular value as a suitable f i e l d test in detecting the more advanced forms of the disease but i s not regarded as s u f f i c i e n t l y sensitive to detect the milder cases. The Hotis test proved d i f f i c u l t to interpret i n many instances. The authors are of the opinion that a study made of the various Hotis reactions might prove of real value i n making this interesting test s t i l l more use-f u l . BIBLIOGRAPHY (1) ROSBLL, J. M. 1955. Laboratory and F i e l d methods of milk testing for the Detection of mastitis. University of Montreal. Contribution Ho. 40, Research, series.. (2) 0ASB, 0. H. 1957. Mastitis from the standpoint of the Practitioner. Journ. Am. Vet. Med. Assn., N.S. Vol. 43, Ho. 2. p.p. 137 - I 5 3 . (3) ROGERS , L.A. 1 9 5 6 . Problems of the F l u i d Milk Ind-ustry. Proc. Ninth Annual Convention State College of Wash. Institute of Dairying p.p. 120 -124. (4) HUOXER, G.J. 1935V The Laboratory Detection of Bovine Mastitis, N. Y. Agr. Exp. Station Bui. 6 2 6 . (5) WILSON, G, S. and associates, 1935. The Bacteriological grading of milk. Privy Council, Med. Research Council. Special Report Series No. 2 0 6 . London, Eng. ( 6 ) HOTIS, R,P, and MILLER; W,T, 1956. A Simple Method for Detecting Mastitis, Streptococci i n Milk.. U^S. Dep. of Agr, CIr, 400. (7) BUCKER, G.J. I957. The Presence of Mastitis Strep-tococci i n Bovine Mammary Tissue. N. Y, Agr. Exp. Station Tech, Bui. 241. (8) DAHLBERG, A. 0,, KUQERA, J. J., HENINGJ.Q. and HUCKER, G. J. 1957. The Composition of Milk as Affected BIBLIOGRAPHY . (continued ) by Latent Mas t i t i s . N. Y.. Agr. Exp'; Station Tech. Bui. 2 39. (9) OLARIC, j?t .0. 1956. The Production of Undesirable . Odours in Haw Milk by Specific Oganisms Associated With Udder Infections. Unpublished. (10) GWATKIN, R. , HADWEN, S., and Le GARD, B. M. 1955. Staphylococcic Infections of the Bovine Udder-Abstracts of Papers Presented at the fourth Annual Ohristman Meeting of the Laboratory Sec. Pub. Health Assoc., Toronto, Dec, 30 and 31, 193.5. (Ll) Me GILVRAY.cO. D.-195.S. Mastitis or Garget i n Gows. Ont. Dept. of Agr. Bui 381. (12) SHAW, A..O,HANSEN, H.O. and NUTTING, R.G. 1957. The R e l i a b i l i t y of Selected Tests for the Detection of M a s t i t i s . Journ. of Dairy Science, Vol. 20, No. 4. p.p. 197 - 203. (15) BLOOD, J . W,+ and ROWLANDS, A. 195,6. The C h l o r i n e Content of .Milk as an Indication of Mastitis,Journ. of Dairy Research, Vol. 7, No. 1. p.p. 47 - 54. 

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