Efficacy of Some Antibacterial on Dromedary Subclinical Mastitis

Efficacy of some Antibacterial on Dromedary Subclinical Mastitis

Haider M. Al-Ramahi Mujeeb T.Shamsa

College of Veterinary Medicine/Babylon University Technical Institute of Al -Najaf.

Abstract

The study was carried out in An Najaf province-Iraq. Two hundred and ninety two milk samples were collected from 73 clinically health she camels, each sample was examined by California Mastitis Test, Somatic Cell Counting, bacterial culturing and antimicrobial susceptibility test. The results showed that the prevalence rate of subclinical mastitis was 17% according to quarter basis and 31% according to animal basis. The mean SCC in CMT negative samples was 2.3×105 cell/ml, while it were 3.5×10 ,2.1 ×10 6 and 4.5×106 in samples of CMT score 1,2 and 3 respectively. The main bacterial isolates was coagulase negative Staphylococci (48%),followed by Staph. aureus (24%),Streptococci(16%) ,Micrococci(6%) and E. coli (6%).The CN-Staphylococci was sensitive to all tested antimicrobials, but antimicrobial resistance was recorded in other isolates include Staph. aureus & Micrococci against Tetracycline and Streptomycin , Streptococci against Gentamicin, Penicillin G and Streptomycin, while the E.coli isolate was resistant to all tested antimicrobials except Gentamicin and Ampicillin.

الخلاصة

أجريت الدراسة في محافظة النجف/العراق ،حيث تم جمع 292 عينة حليب من 73 ناقة سليمة سريريا.تم فحص كل عينة باختبار كاليفورنيا لالتهاب الضرع CMT,عد الخلايا الجسمية SCC والزرع الجرثومي واختبار الحساسية للمضادات الحيوية.

أظهرت النتائج إن نسبة حدوث التهاب الضرع تحت ألسريري كان 17% اعتمادا على عدد الأرباع و31%اعتمادا على عدد الحيوانات المفحوصة.كان معدل عدد الخلايا الجسمية في العينات السالبة لاختبار كاليفورنيا لالتهاب الضرع هو 510×2.3,بينما كان العدد 510×3.5 و610×2.1 و 610× 4.5لدرجات اختبار كاليفورنيا لالتهاب الضرع 1 و2 و3 على التوالي.

كانت العزلة البكتيرية الرئيسية هي Coagulase negative Staphylococci بنسبة48%,تلتها Staphylococcus aureus بنسبة 24% و Streptococci بنسبة 16%واشتركت كل من Micrococci و E. coli بنسبة 6%.

أظهرت عزلة CN-Staphylococci حساسية لكافة المضادات البكتيرية المستخدمة في التجربة بينما أظهرت بقية العزلات مقاومة لبعض هذه المضادات البكتيرية تضمنت مقاومة عزلة Staphylococcus aureus و Micrococci للتتراسايكلين والستربتومايسين وعزلة Streptococci للجنتمايسين وبنسلين جي والستربتومايسين ,آما عزلة E. coli فقد أظهرت مقاومة لكافة المضادات البكتيرية عدا الجنتمايسين والامبسلين.

Introduction

Camels are unique animals which adapted to live in harsh desert conditions, and one of remarkable feature is ability of lactating she-camel to secret 20 litters /day of milk that highly diluted with water for at least 10days with out drinking water (Wernery and Kaaden,2002).According to this fact ,the lactating she-camel play the most important role in surviving of arid population(Al-Sultan,1996).Mastitis is the main important disease of lactating she-camel and cause multiple hazardous effects on animal production as well as human health(Radostits et al.,1994).In past many authors thought that mastitis is uncommon in Camelids (Rahman et al.,1987;Fowler,1989 ),However during the last years many work has been done on mastitis in camel in different camel rearing countries (Barbour et al.,1985;Mostafa et al.,1987 ;Karamy,1990 ;Saad and Thabet ,1993 and Abdurahman et al.,1995).Only one study was performed in Iraq by Al-Ani and Al-Shareefi(1997).The present study was designed to estimate the prevalence of subclinical mastitis in camel in western desert of An Najaf province with attempts to isolate the causative agents and demonstrate their susceptibility to some main antibiotic which used in Iraq.

Materials and methods

The study was carried out in western desert of An Najaf province, from 13April to 22 December2004.Seventy three clinically health, lactating she camel ,reared by local tribes were selected to milk sampling.

Milk-sampling

Out of 73 selected She-camel ,292 milk samples were collected.After teat was swabbed with cotton soaked in 70% ethyl alcohol and discarding the few first milk squirts ,20 Ml of milk were collected in sterile universal bottle and transferring immediately in icebox to laboratory for examination.

California Mastitis Test (CMT )

The CMT was carried out according to procedure described by Schalm et al., (1971),by mixing 2 ml of sample with equal amount of CMT solution in a clean paddle, the reaction was he visually scored as 1 ,2 and 3 depending upon the amount of gel that forms.The reaction was interrupted as follow: Score 1=streak of gel which disappear with swirling ;Score 3= district slime gel formation and Score3= gel develop as convex surface and adhere to bottom of paddle.

Somatic Cell Counting (SCC)

The direct microscopic somatic cell counting DMSCC method ) was carried out by spreading of 1 μ L of thoroughly mixed milk from each CMT positive samples over 1 cm2 area on a glass slides ,which lift to air drying and were stained by Newman-Lampert stain as described by Packard et al. (1992).

Isolation and identification of Bacteria

Bacteriological examinations were carried out following standard methods which described by Quinn et al ,(1994) and Sears et al.,(1993).Briefly ,a loopful of positive CMT milk was streaked on blood agar ,and incubated aerobically at 37 Co for 24-48 hours.Identification of bacterial isolates was made based on colonies morphology ,Gram’s stain reaction, hemolytic characteristics and catalase test.Staphylococci and Micrococci were identified by growth on manitol salt agar,coagulase production,catalase and oxidase tests. Gram’s stain negative isolates were sub-cultured on MacConcky agar and further tested using triple sugar Iron TSI agar ,Indol,methyl red ,voges-proskauer , citrate utilization test, urea and oxidase reaction.

Antibacterial susceptibility test

Antibacreial susceptibility test was performed by disk diffusion method as described by Carter & Cole(1990),by use single antibiotic disks produced by OXOID company include Gentamicin (Genta),Eythromycin (Erythro) ,Tetracycline (Tetra),Ampicilin (Amps) and Cloxacillin(Clox).

Results

A total of 292 quarter milk samples collected from 73 lactating she camels, 50 samples were positive to CMT in percentage rate 17% according to quarter bases and 31% according to animal bases..In other hand ,According to number of affected quarters in same udder, the high prevalence was reported in 2 quarters 43.47%,followed by 1 quarter ,3 quarters and 4 quarter in percentage of 26.09%,17.4% and 13.04% respectively, table 1.

Table 1 Reveal the percentage of affected quarters among 23 animals

The highest percentage CMT score was 2(7.88%),followed by score 3 and 1 in percentage rate of 5.14% and 4.11% respectively ,table 2.

Table 2 Reveal the percentage of MCT scores out of 292 milk samples

Table 3 reveal the results of DMSCC ,the mean somatic cell number in CMT negative samples was 230,000 cell/ml, while it were 350.000 ,2100000 and 4500000 cell /ml in samples of CMT scores 1,2 and 3 respectively.

Table 3 reveal the result of DMSCC according to CMT

The results of bacteriological culturing demonstrated that the highest percentage of isolates was of Coagulase negative Staphylococci 48%,while the percentage of Staphylococcus aureus ,Streptococci ,Micrococci. and E. coli were 40%,16% ,6% and 6% respectively, table 4.

Table 4 Reveal the results of bacterial isolation out of 50 mastitic milk samples

The results of antimicrobial susceptibility test showed that CN Staphylococci was sensitive to all used antimicrobial , the Staphylococcus aureus and Micrococci spp. were resistant to tetracycline and streptomycin , Streptococci was resistant to gentamicin.penicillin G and streptomycin ,while the E.coli was sensitive to gentmicin and ampicilin only ,table 5.

Table 5 reveal the antimicrobial susceptibility of mastitic milk isolates

S: sensitive ; R: resistant

Discussion

Different subclinical mastitis prevalence rates were obtained from studies performed in many camels rearing countries ,such as 45% in Sudan(Bakhiet et al.,1992),81% in Palastine (Guliye et al.,2002),20.5% in India(Mody et al.,1998) ,24% in Iraq (Al-Ani and Al-Shareefi ,1997),28.8% in Ethiopia(Wobit et al,2001) and 66.7% in Kenya (Motofari et al.,2005).These variation may be due to weather and expose of udder to trauma due to ticks or desert plant and anti-suckling devices which used by camel's owner to allow the young calves older than one year are herded together with their dams.All these factors are predispose the udders to bacterial infections (Wobit et al.,2001).

The CMT scores were vary according to severity of inflammation ,type of isolates and lactation stage(Mostafa et al.,1987).In present study the data of CMT scores ,threshold of somatic cells count and its significant correlation to SCC are in agreement with other authors (Al-Ani and Al-Shareefi,1997; Bekele and Molla,2001 and Guliye et al.,2002). The strong positive correlation of CMT with the bacteriological findings indicates that camel milk, like that of cows (Schalm et al.,1971), goats (Ramahi and Al-Nassrawi,2007)sheep (Karamy , 1990), has phagocytic cells that constitute one of the essential defenses against microbial infections. . CMT can be used as a screening test to detect subclinically infected udders of camels (Bekele and Molla,2001 ).

There are divergent opinion as to which bacteria are potentially the primary causal of infectious mastitis (Wernery and Kaaden,2002 ).Present study was in agreement with other studies finding that the Gram's positive bacteria and E. coli are the dominant isolate pathogens of subclinical mastitis ,such as Micrococcus spp.(9),coagulase negative and coagulase positive Staphylococci.(Al-Ani and Al-Shareefi,1997),Streptococci (Bhatt et al.,2004).In other hand, Birhanu et al. ,(2008) considered other dominant pathogen such as Rhodococcus equi , Alcaligens spps , Pasteurella haemolytica , Acinetobacter spps .

The susceptibility of isolates to antibacterial vary from strain to strain and from region to region, therefore ,our obtained data are in agreement with result of Iraqi study(Al-Ani and Al-Shareefi,1997) and disagreement with others (Mody et al. ,1998).

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