Using real-time polymerase chain reaction as an alternative rapid method for enumeration of colony count in live Brucella vaccines

Research (Published online: 08-06-2017)

7. Using real-time polymerase chain reaction as an alternative rapid method for enumeration 

of colony count in live Brucella vaccines

Waleed S. Shell, Mahmoud L. Sayed, A. A. Samy, Ghada Mohamed Al-Sadek, Gina Mohamed Mohamed 

Abd El-Hamid and Abde Hakam M. Ali

Veterinary World, 10(6): 610-615

ABSTRACT

Aim: Brucellosis is a major bacterial zoonosis of global importance affecting a range of animal species and man worldwide. It has economic, public health, and bio-risk importance. Control and prevention of animal brucellosis mainly depend on accurate diagnostic tools and implementation of effective and safe animal vaccination program. There are three types of animal Brucella live vaccines - Brucella melitensis Rev-1 vaccine, Brucella abortus S19, and B. abortus RB51. Evaluation of these vaccines depends mainly on enumeration of Brucella viable count. At present, used colony count method is time consuming, costly and requires especial skills. Hence, the aim of this study is to use and standardize real-time polymerase chain reaction (RT-PCR) as an alternative, quantitative, sensitive, and rapid method to detect the colony count of Brucella in live Brucellavaccine.

Materials and Methods: Four batches of different live Brucella vaccines were evaluated using of conventional bacterial count and RT-quantitative PCR (RT-qPCR) using BSCP31 gene specific primers and probe. Standard curve was generated from DNA template extracted from 10-fold serial dilution of living B. abortus RB51 vaccine to evaluate the sensitivity of RT-qPCR.

Results: Results revealed that three batches of living Brucella vaccines were acceptable for Brucella colony count when traditional bacterial enumeration method was used. Results of RT-qPCR were identical to that of conventional bacterial count.

Conclusion: Results concluded that RT-qPCR was relatively sensitive compared to traditional bacterial colony count of these vaccines.

Keywords: Brucella, colony count, RB51, Rev-1, real-time polymerase chain reaction, S19, vaccines.

Cytological endometritis and its agreement with ultrasound examination in postpartum beef cows

Research (Published online: 07-06-2017)

6. Cytological endometritis and its agreement with ultrasound examination in postpartum beef cows

N. Salah and N. Yimer

Veterinary World, 10(6): 605-609

N. Salah: Department of Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia; Department of Obstetrics and Surgery, Faculty of Veterinary Medicine, University of Diyala, Baquba, 00964, Iraq.

N. Yimer: Department of Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia.

doi: 10.14202/vetworld.2017.605-609

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Article history: Received: 18-02-2017, Accepted: 13-04-2017, Published online: 07-06-2017

Corresponding author: N. Yimer

E-mail: nurdeg2006@gmail.com

Citation: Salah N, Yimer N (2017) Cytological endometritis and its agreement with ultrasound examination in postpartum beef cows, Veterinary World, 10(6): 605-609.

Abstract

Background: Endometritis, which is one of the most common diseases in dairy cows postpartum, causes severe economic losses, including increased open days, calving intervals, and numbers of services to achieve conception.

Aim: This study aimed to evaluate the ultrasound method and its agreement with the endometrium cytology method, which is used to diagnose cytological endometritis in beef cows. Moreover, we determined which method has higher sensitivity and specificity at 4 and 5 weeks postpartum.

Materials and Methods: The study was conducted 20-35 days postpartum. A total of 53 clinically healthy beef cows (28 Brangus and 25 Kedah-Kelantan breeds) from three beef farms were obtained. All cows were evaluated at 4 and 5 weeks postpartum, using ultrasound and cytobrush endometrial examination methods to diagnose cytological endometritis.

Results: Endometrial cytology result showed that 11.3% (6/53) and 9.4% (5/53) of the cows exhibited cytological endometritis 4 and 5 weeks postpartum, respectively. A weak-to-moderate agreement found between the diagnostic methods (k=0.29 - 0.50; p<0.01 and k=0.38 - 0.49) at 4 and 5 weeks postpartum respectively.

Conclusion: The percentage of beef cows that were positive to cytological endometritis was low (polymorphonuclear cells, =8%) at 4 and 5 weeks postpartum. Results showed that the ultrasound method is useful and practical for diagnosing endometritis 4 and 5 weeks postpartum. This method exhibited 60% sensitivity, 93.8% specificity, and a 0.50 kappa value, especially when presence of intrauterine fluids and measurement of cervix diameter used in combination.

Keywords: beef cows, cytology, endometritis, polymorphonuclear cells, ultrasound.

Characterization of hemagglutination activity of emerging Newcastle disease virus in Bangladesh

Research (Published online: 06-06-2017)

5. Characterization of hemagglutination activity of emerging Newcastle disease virus in Bangladesh - Helal Uddin, Kamrul Islam, Mukti Barua, Shariful Islam and Abdul Ahad

International Journal of One Health, 3: 28-35

  doi: 10.14202/IJOH.2017.28-35

Abstract

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Aim: Newcastle disease (ND) is an important viral disease for poultry caused by avian paramyxovirus which can be identified by its nature of agglutination activity with red blood cell (RBC) of different species. The study was aimed to characterize the hemagglutinating (HA) activity of ND virus (NDV) at three different temperatures using RBC of five avian species, six mammalian species, and eight different human blood groups.

Materials and Methods: The study was conducted from January to December 2014 at Chittagong Veterinary and Animal Sciences University. Five avian and six different mammalian species were selected for the study. In each species, two blood samples were collected aseptically. Eight different blood groups (A+, A−, B+, B−, AB+, AB−, O+, and O−) were studied in human. HA test was performed using two virus strains ND lasota and field isolate of very virulent NDV (VVNDV) with mentioned species of RBC at chilling (4°C), incubating (37°C), and room temperature (24°C).

Results: Avian RBC requires less time for agglutination than mammalian RBC. Incubation temperature (37°C) requires lowest time and chilling temperature requires highest time for agglutination of RBC. Duck RBC requires lowest time (17.81 min) while chicken RBC needs highest (57.5 min) time for HA at incubation temperature and at chilling temperature, respectively, against ND lasota virus and with field strain. Goat RBC requires significantly higher time for HA (184.68 min) at chilling temperature than other mammalian species. Human RBC requires almost similar time but O+ and O− blood group do not show any HA activity.

Conclusion: ND is considered as an economically significant disease which is highly contagious in nature infecting many avian species. The threat of ND outbreak to poultry industry necessitates effective control measures to reduce the burden in commercial and backyard farming in Bangladesh.
Keywords: chilling temperature, hemagglutination, incubation temperature, Newcastle disease virus, Newcastle disease virus lasota strain, very virulent Newcastle disease virus strain.

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.