AU622664B2 - Teat dip - Google Patents

Description

OPI DATE 16/10/89 APPLN. I D 3'4352 89 WORL AOJP DATE 09/11/89 PCT NUMBER PCT/AU89/00119 Pcr INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (51) International Patent Classification 4 11) rnagaal Piai ~ubr O8/091 A01N 37/06, A61K 31/20,9/10 (43) terb l ica4 Dt: 5Otbe 99(51,9 A61K 7/50, A61L 2/18 2t)rja ~c, 1 Dt: coe 99(51.9 (21) International Application Number: PCT/AU89/00I 19 (81) Designated States: AT, AT (European patent), AU, BB, BE (European patent), BF (OAPI patent), BG, BJ (22) International Filing Date: 21 March 1989 (21,03,89) (OAPI patent), BR, CF (OAPI patent), CG (OAPI patent), CH, CH (European patent), CM (OAPI patent), DE, DE (European 'patent), DK, Fl, FR (European (31) Priority Application Number: PI 7342 patent), GA ('OAPI patent), GB, GB (European patent), HU, IT (Europeau patent), JP, KP, KR,. LK, (32) Priority Date: 21 March 1988 (2 1.03.88) LU, LU (Europoan patent), MC, MG, ML (OAP[ patent), MR (OAPI patent), MW, NL, NL (European, (33) Priority Country: AU patent), NO, R O, SD, SE, SE (European patent), SN (OAPI patent), SU, TD (OAPI patent), TG (OAPI pa- (71) Applicant (for all designated States except US): DARA- tn) S TECH PTY LTD (AU/AU]; 6th- Floor,-409 -St-,Kilda 'RoadMelbournq, VIC 3004--(AU)- 'zx t J Published 44' 1 y Q N With international search report, (72) Inventor; and Inventor/Applicant (for US only) :KEFFORD, Bruce (AU/AU]; RMB 3085, Rule Road, LardteVC32 nr I (74) Agent; GIBSON, David, Vincent, 29 Sutherland Road, Arniadale, VIC 3143 l 4fL, (54) Title, TEAT DIP (57) Abstract A teat dip composition useful for the treatment of mnastitis in dairy cows, The composition in application f rm Is in the form of an qll-inwater~emulsion, The oil phase includes an maUrae 1at1cdfreape ise i fatty acid, having a carbon Chain length in the range of 016 to

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WO 89/08981 PCT/AU89/001 19 TEAT DIP This invention relates to animal health care products and. in ,particular to a -teat dip for dairy cows, The spread of bacterial infection from cow teats during the mi,1]dng operation results in the spread of the infectious mammtary disease known as rnastitis, The spread of this disease is usual~ly reduced by the use of %cnown bacteriocides such. as chlorine and iodine based compositions, These compositions are usually administered to the teat after removal of the Mil~ing cup by dipping or ppray~ng the teat. It is believed these bacterloc ides kill a substantial numbker of bacter~ia including iastitis pathogens, and thus reducte the spread of bacteria into the mammary gland wher~e mastitis may become evident, There are usually insignificant residual effects of these bacteri ocides between mil$kings. There are problems with the use of these agents including irritation to the teat and teat cracking, To alleviate these problemns# emolient additive$ such as glyceri±ne may be in~cluded in these compositions. HiOWevero even with thei use of these emolionts skin irritation can still occur.

It is an QbjeCt of the present inve'ntion to.

provido a teat dip composition which ajevi--tes; nMe of the problems; of tho prior art teat dips or to provide at le~ast an altOrnatiVa teat dip. composition, 2 Accordingly this invention provides in one form an antimicrobial composition when used as a teat dip comprising an unsaturated fatty acid having a carbon chain length in the range of C16 to C20 having at least 20% by weight of the unsaturated fatty acid with a level of unsaturation of or and a stabilising amount of essentially nonionic surfactant, Preferably the unsaturated fatty acid has a chain length C18 and more preferably the unsaturated fatty acid is linolenic acid.

Preferably the composition is in the form of an oil-in-water emulsion, Th. stabilising amounts of surfactants enable a stable oil-in-water emulsion to be formed, The surfactants include a non-ionic surfactant, in particular a polyethylene oxide derivative, Most preferably the surfactants are substantially non-ionic, Examples of suitable surfactants are sorbitan monolaurate and ethoxylates thereof, Surfactants are selected on the basis of their ability to form stable oil-Hnwater emulsions, Generally it is best if at .'20 least two surfactants are used, one with a HLB value less than 10 and one I with a HLB value in excess of 10. It is important that the surfactants when .used in the teat dip composition do not result in unacceptable residues in milk, The surfactants are used at art recognised levels and are typically of the order 1*20% by weight of oil phase, It Is believed the surfactants also Inhibit the adhesion of mastitis pathogens thereby improving the efficacy of i the composition, The pH of the composition is such that in use irritation to the teat is minimal and a pH in the range 6-8 Is found to he particularly suitable, The relative ratios of oil to water phases are not critical to .30 the working of the composition although from a cost effectiveness viewpoint an oll-n-water ratio in the range of 1/5 to 1/25 is found to be useful in practice, Of course the composition could be packaged for sale at a higher oil-in-water ratio and diluted before use by the dairy farmer by the addition of water, Though this can cause some emulsion stability problems where water quality is poor, Preferably deionfsed water Is used in the compositions of the present Invention, Though less preferred it is possible to sell the oil phase which is emulsified before use by the customer.

The fatty acids suitable for the compositions of the present invention can be selected from fatty acids derived from naturally oecurring tri.glycerde oils such as vegetable tills, for example, linseed, perilla, soya 9 9 9e 0 *9 o* 9 r i t 99..

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9 9 9* rr 9 9i 9 9 *9 99 99« 9. 9 *9 9 9 3 and safflower. Synthetic unsaturated fatty acids may also be used, One particularly suitable active fatty acid is hydrolysed linseed oil. It will be appreciated that with both the synthetic and natural fatty acids, significant range of compositions of fatty acids can occur. Significant proportions of saturated fatty acid may be present. These are not unduly detrimental to the composition, other than diminishing the level of active unsaturated fatty acid in a given oil phase, The unsaturated fatty acid may be selected, for example, from lauroleic, myristoleic, palmitoleic, oleic, ricinoleic, gadoleic, erueic, Preferably the level of unsaturation is or such as in linoleic, linolenic, eleostearic, licanic and arachidic. Other oleofinic components may be present without adversely affecting the working of the invention, However preferably at least 20% or more preferably at least by weight of the composition, or when in the form of a ooilin-water emulsion at least 20% or more preferably at least 30% of the oil phase is 15 unsaturated fatty acid and more preferably at least Compositions of the present invention are found to be effective in reducing the spread of mastitis within a herd of dairy cows and at the same time not increasing teat cracking, It is also noted that residues from the present compositions, in particular unsaturated fatty acids, which may ultimately be introduced into milk are believed to be non-harmful and indeed may be already present at low levels In milk, The present compositions also have a residual effect between milkings that Is greater than the prior art composition.

Other ingredients may be used in the composition such as 25 glycerol, This may be present when hydrolysed vegetable oil is used, Whilst the exact mechanism by which the present composition is effective is unknown tt is believed the unsaturated fatty acid is effective in killing the Staphylococus aureus and Streptococci (gram positive bacterial believed to contribute to mastitis disease control by post milking hygiene, Other bacteria particularly gram negative bacteria are relatively unaffected, Thus some bacterial resistance remains from the unkilled bacteria, For example, harmless bacteria remain as a protective cy*"*

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n~i~ WO 89/08981 PCT/AU89/00119 4 ultimately be introduced into milk are believed to be non-harmful and indeed may be already present at low levels in milk. The present compositions also have 4 residual effect between milkings that is greater than the prior art compositions.

Other ingredients may be used in the composition such as glycerol. This may be present when hydrolysed vegetable oil is used.

Whilst the exact mechanism by which the present composition is effective is unknown it is believed the unsaturated fatty acid is effective in killing the Staphylococus aureus and Streptococci (gram positive bacteria) believed to contribute to mastitis disease control by post milking hygiene.

Other bacteria particularly gram negative bacteria are relatively unaffected, Thus some bacterial resistance remains from the unkilled bacteria. For example, harmless bacteria remain as a protective coating on the teats. This contrasts to the prior art compositions which essentially kill all bacteria, Hence the compositions of the present invention provides some residual bacterial protection between milkings.

The invention will be further described by references to a preferred embodiment in the following Example in which all parts are parts by weight.

Examp3le 1 A teat dip composition was prepared by dissolving: hydrolysed linseed oil fatty acid aeen 20 Span 60 1 Span 60 is added to the hydrolysed linseed oil, heated and stirred until the Span 60 is dissolved, the Tween is then added to this solution, after which this WO 89/08981 PCT/AU89/O01 19 mixture is added to an equal volume of deionised water and agitated. An. emulsion formed which was then diluted with more deionised water to lead to 93 parts of water in the overall composition, The resulting teat dip composition was stable over a period of several weeks with no evidence of the emulsion breaking. The composition was evaluated in vitro by appropriate cell counts of S-aureus bacteria, As c-ontrols water, chlorine and an iodine. based commercial treatments were used, Generally the results showed the composition to be more effective in k~illing the bacteria than either of the commercial coatrol materials, In-vitro testingT of the Composition of Example 1 Additives at various concentr~ations Were added to molten nutrient agar or Todddiewitt agar (for gjrowth of the streptoqocci.) and pl~ates were poured 2G Cultutres of Staphylococcus aureus (1 5 strains from bovine mastitis case..q and Streptococcus agalartae (4 strains f rom bovine mast~tis cases) were spotv,-1 onto tlie plates and these Were examined for, growth afteor 244 at 27'C.

Nweon 20 IS a polysthyltn4 oxide sorbitan monola2rAte, a non-ionic surfactan~t of H-LB 16,7 manufactured by ICI .peciality Chemi~cal Xndpies N.V. Appro4matoly 20 eth~ylene oxide units are corndenaed onto the sorbitan.

AASpAn $0 iS sorbitan, monostoeA~'~e non~-tonic ourfactant of HLB4,1 maaUfaeturod by ICI Spec4tality Chomital Industries NV WO 89/08981 WO 8908981PCT/AU89/001 19 Conc. (ug/rnl) at which no growth was seen on agar plates Additive Staphylococcus Streptococcus aureus acalactiae Untreated 660 660 Active (LSQ)* Active tHLSO)** 330 600 66 Example 1 330 660 66 Surfactants' (Tween 20) 660 660 Surfactants (Span 60) 660 660 In these testas, as in all others, the strains of' StreptococcUs acmalactiae, were mUch more, sensitive to the active ingredient and Example 1 than, the strains of Staphylococcus aureus, Linseed Oil. (LS01 H ~ydrolysed. Linseed, oil fatty acids ti-LSO,) In vivo testing for Intra-mamrnary Infection$ showed that ni~cdozeno cf now tn~'ections (in a her~d of 7S5 COWS determined by bacteri0ologcal CUI~tUre o mastiti's pathoons I~ milk) WASs niricantly les With the OOMpdsition of the Present iWerition. It was AI.to nOted that 'teat Cracking W40 minimal and that the ganorAl Con'dition of the teat4 WAO vdry S06d.

"we WO 89/08981 WO 8908981PCT/AU89/00 119 7 Bacteriocidal Efficiency A teat dip should prove effective in reducing the recoverable number of bacteria (rastitis pathogens) which have been experimentally applied to the teat.

This experiment involved: a. The application of a mastitis pathog'en (Staphylococcus aureus) at a concentration of approximately 5 x 10 CFU/ml to the teats of 10 cows.

b. One front, teat and one rear teat of each izow was treated with the teat dip composition.

C, Untreated teats remained as controls.

d, The number of col~ony forming units of the pathogen recovered by swabbing and culture proced1ures was compared with the number recovered from qontrpl teats, Table I Effect of teat treatment on bacterial count Treatment Mean log 10 decrease in bacterial, numbers Comnposition of Example 1 3.15 Iodine (5foooppm) I .68 Ale 30,a -ClcrhexidineY 1 .51 According to the Draft $tandards an 'feffoctjvell bacteriacide should decreasie thcs number of bctoriA recovered by log 10 1# (or 9P. kill), as These retul.ts show the Compazit~on Of the pro~nt inVeption is Very bacterio id~3ly fCiAent WO089/08981 PCT/AU89/OOI 19 8 and much more efficient than the two prior art teat dips.

Teat Condition This evaluation involved: j a. The application of the teat dip to one front teat and one hind teat at each milking.

b. The remaining teats were untreated or dipped in a registered teat dip.

c. Teats are scored with respect to chaps, sores and. skin condition at fortnightly intervals, d, Comparison of control and test teat scores were made with respect to; total numnber of chapsj (ii) total number of teat soresl (iii) total. teat lesions (ie. chaps plus (tv) numbers of rough skinned~ teata.

Two herds tested in Autumnn (when teat cond~ition is relatively good) showed no significant, Rerults betweea scores treated with the r.omposit iqn according to the invention and the scores of I~odine (5000 ppm) or treated teats, There- 14 no del~eterious effect of the compositi~on of the present tnvention, It is also possible that apositive improvement it teat condition with the coMposition, May be, seen whien the general level of teat condition is poorer, say I Winter, Non-contamination of milk The Use of the teat dip should not impart flavour# odour or colour to m~ilk or mil.k poducts or introduce h~rMtU1 conceratrations,- of uoidues,, W0 8908981 PCT/AU89/OO1 19 9 Results a, Residues It is difficult to test residues of the composition using the standard procedures as there are quantities of the active ingredient in the lipids of normal milk, An estimate of the quantity of this component that could be added to the milk during milking say of the teat dip Is transferred to the milk) indicates that any residue is far outweighed by the natural levels of this compound in the milk lipids.

Estimates: Contribution of active Ingredient to milk =0.25mg/100 mi.

NOTE; This level of the compound is too low ,to detect using current technology, (ii) Content of active ingredient in normal milk~ 20X4 addition, as the other components of the teat dip are FOOD GRADE chemicals at a lower concentration than the Active compound it seems very unlikely that these com-pou~nds cou~ld signif icantly qontamipa-te thea milk under normal conditions, b, ToxIcity for Cheese, Starters and Yoghurt makers The composition of Example I was tested At the cQLl);ert Chandler lfstituto of, Dairy Tochnology' 1 Werribe, for its Off ect on cheese starter and, yo~hurt making bacteria, The effect of test compounds on those bacteria is meaotured In terms of tho change in PHI during the choese Yoghurt making. Zt tho choose containing a, test cornpoiun4, At A given concontrationt has a PH At tho and of the incubation 0,2 units higher than the control. batch thea the toot compound. had A Significant Ofeect, at, that WO089/08981 PCT/AU89OO1 19 concentration, It was found with the composition of Example 1 that at concentrations, 1 ul/mi of milk, there was no significant effect on cheese or yoghurt making.

This concentration (1 ul composition of Example 1/iij of milk) represents a dilution of 1 :1000. For a cow yielding 20 litres of milk 20 ml of composition of Example 1 would be required to signi1ficantly affect cheese and yoghurt making, As approximately 10-15 ml of teat dip is Used per cow, some drains off, and some would be removed during the pre-mllking washing, it is unlikely that under normal milking conditions such a level. of contamination could be achi-eved, Residual Effects When the composition of Example 1 was tested U on the two herds for its effect on teat skin, sWalg were also taken of the tOea that had been treated ZQ twice daily With either the com(psition of Example 1, iodine (5000 ppnM) or Untreated for 8 week~s, At thje end of, the experiment swabs, of all. teats were -taken 14 hours; af ter troAtiijent, and serial dilutions of fluid from, them were plated onto staphyltococcal selective (sheep blood agar) Two bacterial types groew on the selectiva medium, (staphylococci, And b~oillll whereas a range of, colony types inluding the above were seen on the non-selectiVoe plates "total" bacteria).

Swabs from-l the com~position of Exap~le I treated toats whon Oompoirad to iodine-trotd1 teats hAvqt 2. timos fawdw1 ztAphyec& I: seoutivo I's, tim n2,ca lzacil2l I mdltiw' WO 'W89/08981 PCT/AU89/OO1 19 and 1 .5 times fewer "total" bacteria )non-selective me di um Swabs from the composition of Example 1. treated teats i~ when compared with untreated teats have; j 4,6 times fewer staphylococci 1,A times more bacilli and 4,5 times fewer "total" bacteria results suggest: The composition of Example 1 is more effective than iodine in holding staphylococcal numbers down between mllXing4, (iil) The composition, of Example 1 promotes the of bacill (and possibly other l~ess pathogenic bacteria on teats which may lead to reduced reinfection of the teat through ;omnpetition, (iII) Composition, of, Example 1 reduced: the "total" populato of bacteria on teat.

Since modif icationg within, the spirit and scope of the invention may be readily eff~ected by prsoas zii3Jed in the art, It is to be under'stod thAt Cho invention is not limited to thle particular ombodiiint described, by Way of example, hereinabove#

Claims (3)

1. 2. An anti~microbial teat dip composition when used as a teat dip as defined in Claim I wherein the composition is5 in the form of an ollln~wpter emulsion,
2. 3. A, teat dip composition, as defined in Claim 1 or 2 wherein at least 20% by weight of the unsatUrated fatty acid is linseed oil fatty acid, 4, A teat dip composition as defined in Claim 3wherein the rnsaturated. fatty acid' is %9 linseed oil fatty goid 61 A teot dip composition, as defined in Claim 4 further Including glycerol,
3. 6. A toot dip composition as dqfined in Claim 5 wherein there are. at least two, $Lrfacants, i 0one with HMB Ims than 10 and. one, with HL, greater than 44 4 _o I I ERNATIONAL SEARCH REPC T International AppLication No, T=/AU 89/00119 1 1. CLASSIFICATIONl OF SUBJBT MATTER~ (it several classification symbols apply, indicate a)6 According to International Patent Classification (IPC) or to both National Classification and IPC nt Cl. A01N 37/06,A61K 31/20, 9/10, 7/50 A61L 2/18 1 Minimum Documentation Searched 7 CLassification System IClassification SymboLs A61K 31/20,9/10 A61L 2/18 AO1N 371/06 Documentation Searched other than minimum Documentation to the Extent that such Documents are Included in the FieLds Searched 8I AW, nr as above III, raJMumS COMIDRa TO BE Rd=ANT 9 catqgory* citation of Document, i;with indication, where appropriate, I ReLevant to I of the relevant passages 12 jClaim No 13 I Xt Cosmetic arud Drug Preservation 1984 Jckw= J Kabara. (13-7) 'Mdindan. Fatty Acids and Esters as Antimicrobial Agents, jI see pages 275-304, especially pages 279-283, I Y AU,A, 71803/87 (-ABA1RA) 22 0Qtober 1987 (22.10.87) see pages 3, (1-7) I I 313-14 and claims. Y AU,A, 71804/87 (KABARA) 22 October 1987 (22.103.8) See pages 3 and I(1-7) claims. I XY WO,A, 84/02Z71, MHAS T) 21. Jurp 194 (21,06,84) See pages 5-6 and (1-6) claims !I X AIJ,A, 62696/86, (AIEMICAN CYAMMI COMIA) 19 Mar 1987 (3.9.03.87) -37 See claims 1-2, I (continued~)I 4 pociak categories of Cited documentsi 10 Laterdcmn qj~~e fe the I intornationa. Ming date or priority doteL I A, document defining the generak state of the and not in confLict with the apptiqatlon bt I art which is not considered to be of OWte to understand the principko or theory particulamr rotovance underLying the invention IV earlier document but pub~cIahed on or document o( partiduLar re~ovahce; the I after the InternationaL filing date, claimed invention cannot be, cqnslderecl novel, document which may throw4 doubtt on priority or cannot be 4onsidered to invo~ve On claim(s) or which is cited to ostAtalsh the inventive step I I publication date of arnother citation or document of part~cfkar, ro~oVancqt the I other, special; reasont (as specifled) c aimqd Invention tannot be considered t4 document referring2 to an orak I~oir. involve an inventive step when the document I uso, oxhibition or other, means is Combined with onle Of' more other su4h IP 004doumqnt Published priorf to the do4umants, ;uch 0.ombination, being Obvious to intoriiatlonel Miing clae, taut Waer thani a Person ski~ld in the art, I the priority date c~aiued *91 document member of the same patent farsiLy IDate of the Actua \ompketion of the I Date of Maiifg of thils JnternationaL t nternationst $oorch I qemrch Report[ 1$ Jmw 139 (MMW",O J L I nternationak. qArchlng Authority igture Of Au(fo$Ijd Officer Aus 4 raun 5 I)t~ft OMkC4 Form PCT11SAY211Q (socond sheet) (Januafy 1948 I U international AppL adtion No. MrZ/AU 89/00119 RMTHM IaMIMON CONTMM FEM IM SOMM M= I X iAIJB, 17589/83 (559715) (TME REMTS OF M~1 MaNVUSITY OF (1-7) I I CALIFORIA) 16 February 1984 (16.02.84) See page 4 and claims 8-12. X AUSB, 38967/78 (525236) (BFMiJ Er AL) 21. February 1980 (21.02.80) I(1-2,7) See especially pages 7-8 and claim 1. I X AIJ,B, 19947/83 (567890) (CIBA-MIGz') 12 April. 1984 (32.04.84) j(1-2,7) See whole docuzmmt. I x US,A, 3949071 (91Ml~) 6 April 1976 (06.04.76) See colums 4-5. I(1,7) A JAXJ,B, 14932/83 (55 (VILSON4) 29 Novwber 1984 (29-11.84) I(1,7) See whole docuumxt is intrainLvac eothsnt enatbihdi epc fcrancsm e I C I 172(* o thet~oigraos I ~im nubr beas hycIt osbec atrntrqie ob serce by thi Auhrt, mL- Z ,C3 Ckaim numbers because they relate to parts of the internationl, appLication that do I comply wth the prescribed requirement; to such an extent that no meaningful international I search can be carried, out, specifcaktY-. 3,[3 ;kai* numbers because, they are dependent Claims and are not drafted in accordance with the second and third sentences of PCT Wue 6,4 (a)t VT. 3 BSEVA'IM1 WM~. UflM OF MOrxcN IS LCK 2 IThis ;ftternationaL Searching Authority found multiple inventions in this internationaa ppLicatn as falkLaws. i 1""As aWl r. 8eoa itionaL search fees were timely paid by the arpicant, this intern~tionaL Isearch, report coves Wl searchable otafms Of the! Internationa, application. Ia~ At onky too* of the required ditiona, search fees were timely paid by the app(icant, this Iinternational search report cover* only' those O(Oims of the internation4l apptication for Iwhich fees were, paidi- speoiftcaLy 4laimst I3,C)NQ- required additionat- searth feos were timely paid4 by the aopicnt. Consequenty, this Iinktnationat search report is restricted to the inyvtioti first mentioned in the ckeaut Iit It qovored by gtlaim umhers C! r At, al(1 searchable ckaeut oottf be searched without effort justifying an additional fee, Ithe Itlrnatianat Swac*hino Authority did not invite, payment Of any additrionl, tee, IRemark on Protqst tI C he 4ddition~t soardh feos; Wert Ocompontecl by applicane's protest, El0 No protest accompanted the payment of addii ona( search, feet. forma PCT/I$A/210 (sopplemental sheet (Jonuary tMaS