Molecular characterization of antibiotic-resistant Staphylococcus aureus from livestock (bovine and swine)

Research (Published online: 05-06-2017)

5. Molecular characterization of antibiotic-resistant Staphylococcus aureus from livestock 

(bovine and swine)

Asima Zehra, Randhir Singh, Simranpreet Kaur and J. P. S. Gill

Veterinary World, 10(6): 598-604

Aim: The aim of this study was to figure the prevalence, phenotypic and genotypic antibiotic resistance (AR) pattern of Staphylococcus aureus isolated from bovine and swine nares.

Materials and Methods: Colonies with typical morphology on Baird-Parker agar supplemented with egg-yolk tellurite emulsion were selected and biochemically/genotypically identified as S. aureus. These strains were further subjected to epsilometer test for their sensitivity to various clinically important antibiotics and antibiotic susceptibility testing for amoxicillin/clavulanic acid, and double-disk diffusion testing was performed by the standard disc diffusion method following CLSI guidelines. S. aureus strains were also tested for the presence of AR genes, viz., blaZ, mecA, aacA-aphD, 

erm (ermA, ermB, ermC), tet (efflux genes tetK and tetL, tetM and tetO of the ribosomal protection family), and vanA.

Results: The nasal cavities of 17 out of 47 randomly selected bovine and 20 out of 28 randomly selected swine were positive for S. aureus, representing the prevalence of 36.2% (95% confidence interval [CI]: 22.5-49.9) and 71.4% (95% CI: 54.7-88.1), respectively. Most of the S. aureus strains showed higher resistance to penicillin (94.6%, minimal inhibitory concentration [MIC] =1.5 μg/ml) followed by ciprofloxacin (56.7%, MIC =32 μg/ml) and tetracycline (18.9%, MIC =32 μg/ml). About 10-15% of the strains were resistant to gentamicin (MIC 16 μg/ml) and oxacillin (MIC 6-8 μg/ml). None of the strains were resistant to vancomycin (MIC 0.25-1.5 μg/ml). In this study, 32.4% strains were resistant to three or more than three antibiotics and prevalence of this multi-drug resistant S. aureus 

was 45% (95% CI: 26.6-63.4) and 17.6% (95% CI: 6.7- 28.5) in swine and bovine nasal samples, respectively. Four strains from pigs were borderline oxacillin-resistant S. aureus MIC 6-8 μg/ml, but none were mecA positive. Two of these strains were β-lactamase hyperproducers. Among the resistance genes blaZ, tetK, tetL, tetM, ermB, and aacA-aphD were found.

Conclusion: Our results demonstrated the absence of mecA and pvl gene, but the presence of multi-drug resistant S. aureus in the nares of healthy animals which has a potential to spread in a community.

Keywords: antibiotic resistance genes, epsilometer test, livestock nasal swabs, multidrug resistance, Staphylococcus aureus.

Prevalence of methicillin-resistant Staphylococcus aureus skin and nasal carriage isolates from bovines and its antibiogram

Research (Published online: 04-06-2017)

4. Prevalence of methicillin-resistant Staphylococcus aureus skin and nasal carriage 
isolates from bovines and its antibiogram

Alok Kumar, Purushottam Kaushik, Anjay, Pankaj Kumar and Manoj Kumar

Veterinary World, 10(6): 593-597

Aim: This study was conducted to determine the prevalence of methicillin-resistant 
Staphylococcus aureus (MRSA) in cattle and buffalo and to study their antibiotic 
resistance pattern.

Materials and Methods: A total of 136 samples (skin and nasal swab) from cattle and buffalo 
were collected. MRSA was identified by conventional bacterial culture techniques which were 
further confirmed by amplification of S. aureus-specific 16S rRNA by polymerase chain reaction 
(PCR). The isolates were further analyzed for the presence of mecA gene by PCR. The 
antimicrobial susceptibility profiling was performed by disc diffusion method.

Results: The prevalence of MRSA in the current study was 28.57% and 34.28% in cattle nasal 
and skin swab, respectively, with an overall prevalence of 31.43% MRSA among cattle. Buffalo 
nasal and skin sample showed MRSA prevalence of 54.55% and 39.4%, respectively, with 46.9% 
overall prevalence. PCR could detect mecA gene in 36.4% and 58% MRSA isolates from cattle 
and buffalo, respectively. Antimicrobial susceptibility test found MRSA resistant to penicillin and 
oxytetracycline (88% each), cefoxitin (75%), cotrimoxazole (62%), and amoxyclav (50%). 
100% sensitivity was observed against ciprofloxacin, amikacin, chloramphenicol, and gentamicin. 
Three (16.7%) MRSA isolates from buffalo were found resistant to vancomycin.

Conclusion: Cattle and buffalo were identified as a potential carrier of MRSA in Bihar (India). 

The isolation of vancomycinresistant S. aureus (VRSA) in the current study indicates the 
emergence of VRSA in animal population which may be transmitted to the human beings 
working in close contact to the animals.

Keywords: antibiogram, bovine, mecA gene, methicillin resistant Staphylococcus aureus.

Traditional treatment of human and animal salmonelloses in Southern Benin: Knowledge of farmers and traditherapists

Research (Published online: 03-06-2017)

3. Traditional treatment of human and animal salmonelloses in Southern Benin: Knowledge of farmers and traditherapists

T. V. Dougnon, E. Deguenon, L. Fah, B. Legba, Y. M. G. Hounmanou, J. Agbankpe, A. Amadou, H. Koudokpon,  K. Fabiyi, A. Aniambossou, P. Assogba, E. Hounsa, M. de Souza, F. Avlessi, T. J. Dougnon, F. Gbaguidi, M. Boko, H. S. Bankole and L. Baba-Moussa

Veterinary World, 10(6): 580-592

ABSTRACT

Aim: This study aimed to report medicinal plants that are likely to be used in the control of salmonellosis.

Materials and Methods: A cross-sectional study was conducted in Southern Benin. Semi-structured questionnaires were administered

to 150 farmers and 100 traditional therapists in seven high municipalities. This step helped to collect plants that are used in the treatment

of animal salmonellosis and typhoid fever in human.

Results: The results revealed a low level of use of medicinal plants among breeders who prefer antibiotics such as oxytetracycline (53.55%),

tylosine + sulfadimerazine (15.30%), and alphaceryl (19.13%). However, plants such as Moringa oleifera (leaves), Carica papaya 

(leaves and seeds), and Vernonia amygdalina (leaves) were mostly used by some farmers. From traditional therapists, 57 plant species of

32 families were identified as typhoid fever cures; among which Leguminosae, Asteraceae, and Euphorbiaceae were predominant. 

Persea americana (22.72%), V. amygdalina (7.57%), and Corchorus olitorius (7.57%) were the most cited by traditherapists for the

treatment of typhoid fever in human.

Conclusion: This study provides a database for further studies on the in vitro and in vivo efficacy of Benin plant species on 

Salmonella spp. These evaluations will guarantee the availability of new therapeutic solutions for populations.

Keywords: farmers, medicinal plants, salmonellosis, Southern Benin, traditherapists.

Prevalence of paratuberculosis in organized and unorganized dairy cattle herds in West Bengal, India

Research (Published online: 02-06-2017)

2. Prevalence of paratuberculosis in organized and unorganized dairy cattle herds in

West Bengal, India

Jitendrakumar M. Bhutediya, Premanshu Dandapat, Arijit Chakrabarty, Ratan Das, Pramod Kumar

Nanda, Samiran Bandyopadhyay and Tapas Kumar Biswas

Veterinary World, 10(6): 574-579

ABASTRACT

Aim: The aim of this study was to determine the prevalence pattern of Mycobacterium avium subsp. paratuberculosis, the causative

agent of paratuberculosis or Johne's disease, in unorganized as well as organized cattle herds in West Bengal.

Materials and Methods: Four organized cattle farms with identical management practice in Nadia (n=3) and South 24 Parganas (n=1)

districts and three unorganized cattle herds, one each from three districts, namely, Burdwan, North 24 Parganas, and Purba Midnapur,

were selected randomly and screened for paratuberculosis by delayed-type hypersensitivity (DTH) and enzyme-linked immunosorbent assay (ELISA).

Results: Of 191 animals tested by DTH, 57 (29.8%) were found to be positive in comparison to 72 (37.7%) by ELISA. In organized farms,

seropositivity varied from 13.3% to 53.1%, whereas in unorganized sector, it ranged from 5% to 6.7% with one area having exceptionally

high prevalence, i.e. 53.3%. The range of positivity detected by DTH both in organized farms and backyard sectors varied from 0% to 46.7%.

By employing both DTH and ELISA together, the positivity of animals in organized and unorganized herds was 19.9% and 8%, respectively.

Conclusion: The results indicate that animals in organized farms are much more prone to paratuberculosis than others.

For screening the herd, both DTH and ELISA should be used simultaneously to increase the test sensitivity in order to minimize its further

spread adopting control programs.

Keywords: cattle, India, Johne's disease, paratuberculosis, prevalence, West Bengal.

Efficacy of early chick nutrition with Aloe vera and Azadirachta indica on gut health and histomorphometry in chicks

Research (Published online: 01-06-2017)

1. Efficacy of early chick nutrition with Aloe vera and Azadirachta indica on gut health and histomorphometry in chicks

Tamilvanan Sujatha, Sivasankar Abhinaya, Jai Sunder, Marudhai Thangapandian and Anandamoy Kundu

Veterinary World, 10(5): 569-573

Aim: This study was conducted with an aim of studying the efficacy of water supplements of Aloe vera and Azadirachta indica (neem) during pre-starter

age (0-2 weeks) on gut health and histomorphometry in Vanaraja chicks.

Materials and Methods: A total of 192 day old Vanaraja chicks were randomly assigned to one of four 

herbal water treatments throughout the experimental pre-starter stage (0-2 weeks) in a completely 

randomized design. Each treatment was given four replicates consisting of 12 chicks per replicate. Water treatments comprised T1: Control with regular antibiotic supplement, T2: 3 ml Aloe juice per chick per day, 

T3: 3 ml neem extract per chick per day, T4: 1.5 ml Aloe and 1.5 ml neem per chick per day. Gut culture 

was done for Escherichia coli and Lactobacillus sps. and gut histomorphometry in 24 gut samples at 14 

days of age.

Results: This study revealed that supplementation of A. vera and neem in water significantly (p<0.05) 

reduced and increased the number of gut E. coli and Lactobacillus sps. Colonies, respectively, as 

compared to that of control groups; Villi was significantly (p<0.05) taller and broader on 14 days of age

across the jejunum of chicks fed with neem supplementation as compared to that of control chicks. 

Significantly lower crypt depth (p<0.05) was observed in the duodenum of Aloe supplementation. 

Villus height: Crypt depth ratio of duodenum and jejunum was significantly (p<0.05) increased neem and 

Aloe supplementation in chicks as compared to their combination and control.

Conclusion: Immediate post hatch supplementation of Aloe juice and neem extract in chicks improved the development and health of their gut.

Keywords: Aloe vera, Azadirachta indica, chicks, early chick feeding, gut health, histomorphometry.

Immunopathological response of leukocytes against microfilariae and adult worms in white rats infected with Setaria cervi

Research (Published online: 30-05-2017)

16. Immunopathological response of leukocytes against microfilariae and adult 
worms in white rats infected with Setaria cervi

Sharba Kausar and Wajihullah Khan

Veterinary World, 10(5): 562-568

ABSTRACT

Aim: Aim of this study was to see the immunopathological changes against the microfilariae (Mf) and adult worms of a bovine filarid, Setaria cervi in the tissues of vital organs of experimentally infected white rats. The effect of diethylcarbamazine (DEC) was also observed on the Mf, as leukocytes especially lymphocytes produce immunoglobulins which opsonize and increase the efficacy of DEC against circulating Mf. Effect of this drug was also assessed on liver enzymes in the microfilaremic rats.

Materials and Methods: Microfilaremia was established by implanting adult worms intraperitoneally and by the infusion of Mf recovered from the uterus of gravid female worms. DEC was administered orally for six consecutive days in the rats having patent infection. Differential leukocyte count was recorded every 3rd day, and liver enzymes were estimated every 10th day in both treated and untreated rats. Pathological changes were observed in HE stained sections of vital organs where Mf or adult worms were trapped.

Results: Destruction and reduction in microfilarial density were noticed in microfilaremic rats treated with DEC. Trapped Mf and embedded worms revealed heavy cellular infiltrations by defensive cells which surrounded and attached with the body surface of the Mf as well as adult worms for their destruction and piece meal clearance. Immune-mediated pathology was observed in the tissue sections of lungs, spleen, and liver. Liver enzymes were elevated during the period of higher parasitemia.

Conclusion: There was a moderate level of immunopathology against the Mf and adult worms by the leukocytes in experimentally infected microfilaremic rats. Mf were in the process of degeneration where they got trapped. Moderate increase in liver enzyme was noticed which was slightly more in untreated group. Although a fraction of Mf gets killed in the peritoneum, majority of them successfully enter the systemic circulation and survive for about 54 days, which is sufficient enough for conducting immunological and chemotherapeutic studies.

Keywords: differential leukocyte count, liver enzymes, microfilariae, pathology, Setaria cervi, white rats.

Prevalence of multiple drug resistant Streptococcus suis in and around Guwahati, India

Research (Published online: 28-05-2017)

15. Prevalence of multiple drug resistant Streptococcus suis in and around Guwahati, India

Mrinalee Devi, Jyoti B. Dutta, Swaraj Rajkhowa, Dhireswar Kalita, Girindra Kumar Saikia, Bipin Chandra Das,

Razibuddin Ahmed Hazarika and Gauranga Mahato

Veterinary World, 10(5): 556-561

ABSTRACT

Aim: This study was conducted to determine the prevalence and antimicrobial susceptibility of Streptococcus suis and their resistance patterns isolated from both clinically healthy carriers and diseased pigs in and around Guwahati, Assam, India.

Materials and Methods: A total of 497 samples were collected during October, 2012, to April, 2014, from clinically healthy (n=67) and diseased (n=230) pigs of varying age and either sex maintained under organized and unorganized farming systems. Samples were processed for isolation and identification of S. suis by biochemical characterization and polymerase chain reaction targeting the housekeeping gene glutamate dehydrogenase. In vitro antimicrobial susceptibility of the recovered isolates against nine antibiotic groups comprising 17 antimicrobial agents was studied by standard method.

Results: Of the 497 samples examined, 7 (1.41%) isolates were confirmed to be S. suis of which 5 (1.87%) and 2 (0.87%) were derived from clinically healthy and diseased pigs, respectively. All the isolates were susceptible to gentamicin, amikacin, and erythromycin (100%) followed by the penicillin group and enrofloxacin (85.71%), ceftriaxone, doxycycline HCL, ofloxacin and chloramphenicol (71.43%), to kanamycin, clindamycin and co-trimoxazole (42.85%). The isolates showed least susceptibility to cefalexin, tetracycline and streptomycin (28.57%). All the five S. suis isolates from clinically healthy pigs were susceptible to penicillin G, amoxyclav, doxycycline HCl, gentamicin, amikacin and erythromycin, 80.00% isolates susceptible to ampicillin, enrofloxacin and ofloxacin, 60.00% to ceftriaxone, kanamycin and chloramphenicol, 40% to cefalexin, tetracycline, clindamycin and co-trimoxazole, respectively. Only 20.00% isolates were susceptible to streptomycin. Both the isolates recovered from diseased pigs were susceptible to ampicillin, ceftriaxone, gentamicin, amikacin, enrofloxacin, erythromycin, and clindamycin. On the other hand, both the isolates were resistant to cefalexin, tetracycline, doxycycline HCL, and kanamycin. Altogether five different resistance patterns (multi-drug resistance) were observed. Of the seven S. suis isolates, two isolates were susceptible to all the 17 antimicrobial agents, one isolate was resistant to four antimicrobial agents, two isolates to seven agents, one isolate to nine agents, and one isolate exhibited resistance to 14 antimicrobial agents.

Conclusion: This study was conducted to determine the prevalence of S. suis in clinically healthy and diseased pigs and their antimicrobial susceptibility patterns. All the isolates were susceptible to gentamicin, amikacin and erythromycin, and most of them were resistant to cefalexin, tetracycline and streptomycin. Five different patterns of antimicrobial resistance (multi-drug resistance) were observed.

Keywords: antimicrobial resistance, emerging zoonotic pathogen, multiple drug resistance, pathogen prevalence, Streptococcus suis.