Identification and characterization of a novel infectious bursal disease virus from outbreaks in Maharashtra Province of India

Research (Published online: 29-10-2018)

25. Identification and characterization of a novel infectious bursal disease virus from outbreaks in Maharashtra Province of India

Sudhakar P. Awandkar, Prabhakar A. Tembhurne, Jeevan A. Kesharkar, Nitin V. Kurkure, Sandeep P. Chaudhari, Sachin W. Bonde and Vijay C. Ingle

Veterinary World, 11(10): 1516-1525

ABSTRACT

Aim: The study was undertaken to isolate infectious bursal disease virus (IBDV) from clinical cases in broiler and cockerel flocks of Maharashtra state, India, and its molecular epidemiological investigation.

Materials and Methods: The morbid bursal tissues were collected from flocks suspected for IBD. The samples were subjected for virus adaptation in primary chicken embryo fibroblast (CEF) cells followed by confirmation by reverse transcription polymerase chain reaction (RT-PCR) for partial VP2 sequence and phylogenetic analysis.

Results: The isolation of IBDV from field samples took seven blind passages for adaptation in CEF. The cytopathic effects included rounding, aggregation, vacuolation, and detachment of the cells. The RT-PCR showed amplification of 627 bp amplicon specific to the primers for VP2 gene fragment which confirmed successful adaptation and isolation of IBDV using CEF. The nucleotide and deduced amino acids based on phylogeny clustered the current isolate in a distinct clade with classical virulent and antigenic variants. It showed divergence from very virulent (vv) and vaccine strains of Indian origin. The isolate showed unique amino acid substitution at A329V as compared to all other IBDVs. The variation in key amino acids was reported at A222, I242, Q249, Q253, A256, T270, N279, T284, I286, L294, N299, and V329. It shared conserved amino acids at position A222, I242, and Q253 as reported in vvIBDV isolates. However, the amino acids reported at position T270, N279, T284, L294, and N299 are conserved in classic, antigenic variant and attenuated strains of IBDV. The amino acids at positions N279 and T284 indicated that the isolate has key amino acids for cell culture replication.

Conclusion: The IBDV field isolate does not reveal the full nucleotide sequence signature of vvIBDV as well as vaccine strains. Hence, we can conclude that it might not belong to vvIBDVs of Indian origin and the vaccine strain used in the region. This may be suggestive of the evolution of the IBDV in the field due to the coexistence of circulating field strains and live attenuated hot strains, resulting into morbidity and mortality, warranting the need for safer protective vaccines, and implementation of stringent biosecurity measures to minimize loss to farmers.

Keywords: adaptation, chicken embryo fibroblast, epidemiology, isolate infectious bursal disease virus, VP2.

Genetic resistance of eight native Egyptian chicken breeds having chicken B-cell marker 6 gene post-challenge with field strain of Marek's disease-induced tumor virus

Research (Published online: 29-10-2018)

24. Genetic resistance of eight native Egyptian chicken breeds having chicken B-cell marker 6 gene post-challenge with field strain of Marek's disease-induced tumor virus

Hala A. Shaheen, H. A. Hussein, M. M. Elsafty and M. A. Shalaby

Veterinary World, 11(10): 1510-1515

ABSTRACT

Aim: The aim of this work was to detect chicken B-cell marker 6 (ChB6) gene in some native breeds in Egypt and find the relationship between founded genes in these different breeds to determine the resistance of native Egyptian breeds of chicken to Marek's disease (MD).

Materials and Methods: A total of 14 different chicken breeds (30 each) including ten native breeds in addition to SPF Lohmann, High Line, Bovans, and Roodiland were used. Blood samples were collected for the detection of (ChB6,) by polymerase chain reaction (PCR) assay and sequenced to determine the presence or absence of ChB6 gene. Experimental infection was done using local field isolated MD virus (MDV) of 11 (1 day old) unvaccinated chick breeds having no maternal antibodies against MDV. Ten breeds of them carry ChB6 gene, eight breeds were native, and the rest two breeds were SPF Lohmann and High Line in addition to a group of ChB6 gene-lacking breed (Bovans) were infected. Spleen samples were collected from all infected breeds at 20th, 25th, 30th, 35th, and 40th weeks post-infection and tested by PCR assay for the detection of MDV. Furthermore, at 40th week post-infection, tumorized spleen sample of Bovans breed was collected and prepared for examination by transmission electron microscope (TEM) to confirm the presence of MDV.

Results: Our results revealed the positivity of 10 out of 14 breeds (Gimmizah , Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) to the presence of ChB6 gene and resistance to MDV infection, while the Bovans, Mandarah , Inshas and Roodiland breeds lack the ChB6 gene and are susceptible to MDV infection. The collected spleen samples revealed negative for the presence of challenged MDV by PCR in 10 breeds (Gimmizah, Sinai, Dandarawi, Fayoumi, Golden Montazah, Matrouh, Beheri, Dokki, SPF Lohmann, and High Line) and positive for Bovans breed. TEM is used to confirm MDV infection in Bovans group which demonstrated tumors.

Conclusion: The study confirms the relationship between the presence of ChB6 gene in our native breeds and the absence of tumors.

Keywords: ChB6 gene, Egyptian chicken breeds, Marek's disease, polymerase chain reaction, transmission electron microscope.

Bacteriological survey of bulk tank milk from dairy farms in Montero, Santa Cruz, Bolivia

Research (Published online: 26-10-2018)

23. Bacteriological survey of bulk tank milk from dairy farms in Montero, Santa Cruz, Bolivia

Yu Michimuko-Nagahara, Yu Tamura and Masateru Koiwa

Veterinary World, 11(10): 1506-1509

ABSTRACT

Background and Aim: Recently, bacterial surveys for mastitis-causing pathogens in bulk tank milk (BTM) have been conducted in several countries worldwide. However, no such surveys have been reported from Bolivia. Therefore, the present study aimed to estimate the prevalence of mastitis pathogens in BTM from dairy farms in Montero, Santa Cruz, Bolivia.

Materials and Methods: Between July 2016 and August 2017, a total of 43 BTM samples were collected from 3264 cows to determine bulk tank somatic cell counts (BTSCC) and identify mastitis-causing bacteria. BTSCC was classified as follows: = <100×103, 100-500×103, 500-1000×103, and >1000×103 cells/mL.

Results: Mastitis-causing pathogens identified by agar medium cultures included Bacillus spp., coagulase-negative staphylococci (CNS), coliforms, Staphylococcus aureus (SA), streptococci, and other species. The proportions of BTSCC of <100×103, 200-500×103, 500-1000×103, and >1000×103 cells/ml were 0%, 37%, 51%, and 12%, respectively. The proportions of coliforms, streptococci, CNS, Bacillus spp., SA, and others detected in BTM were 33%, 30%, 16%, 7%, 2%, and 16%, respectively.

Conclusion: Although the herd prevalence of contagious mastitis-causing pathogens, such as SA, in BTM was low, increased BTSCC were identified in Montero, Santa Cruz, Bolivia.

Keywords: bacterial survey, Bolivia, bulk tank milk, dairy farm, mastitis.

The effect of Toxoplasma gondii on plasma serotonin concentration in sheep

Research (Published online: 26-10-2018)

22. The effect of Toxoplasma gondii on plasma serotonin concentration in sheep

Annamaria Castello, Giuseppe Bruschetta, Renato Paolo Giunta, Anna Maria Fausta Marino and Alida Maria Ferlazzo

Veterinary World, 11(10): 1500-1505

ABSTRACT

Background and Aim: Toxoplasma gondii is an intracellular parasite that commonly infects warm-blooded animals, including humans. Virtually all species can be infected, but a species-specific variability is evident, in terms of both type and severity of the symptoms encountered. As serotonin (5-hydroxytryptamine [5-HT]) plays an important regulatory role in both physiological and immune responses, the aim of this research was to assess whether toxoplasmosis disease could affect plasma 5-HT concentration and/or hematochemical parameters in a particularly susceptible species to infection as sheep.

Materials and Methods: 5-HT plasma levels were analyzed in platelet-poor plasma fraction by enzyme-linked immunosorbent assay. Blood count and hematochemical parameters were evaluated. Total proteins (TPs), glucose (Glu), and lactate dehydrogenase were determined by a spectrophotometer.

Results: Results showed significantly higher levels in plasma 5-HT, monocytes, and TP and significantly lower levels of Glu, in infected sheep compared to the control group.

Conclusion: Results could support the hypothesis of an effect of toxoplasmosis infection on plasma 5-HT concentrations in sheep. More research is needed to assess the function of 5-HT in the regulation of infected sheep's immune responses.

Keywords: monocytes, plasma, serotonin, serum, sheep, toxoplasmosis.

Evaluation of antioxidant, diuretic, and wound healing effect of Tulkarm honey and its effect on kidney function in rats

Research (Published online: 25-10-2018)

21. Evaluation of antioxidant, diuretic, and wound healing effect of Tulkarm honey and its effect on kidney function in rats

Hamada Imtara, Noori Al-Waili, Meryem Bakour, Wail Al-Waili and Badiaa Lyoussi

Veterinary World, 11(10): 1491-1499

ABSTRACT

Aim: The composition and activity of honey depend on its floral origin. Honey collected from Tulkarm was evaluated for physicochemical property and antioxidant content as well as a diuretic and wound healing activity. Its effect on kidney function was evaluated and compared with furosemide.

Materials and Methods: Honey was collected in Tulkarm, Palestine, and its phenol, flavones, and flavonol content were assessed. The antioxidant activity was determined with the use of colorimetric assays, 1,1-diphenyl-2-picrylhydrazyl, ferric reducing antioxidant power, and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid). Two sets of experiments were conducted. First experiment: 18 rats were used for the evaluation of diuretic activity of honey. The rats received either honey or furosemide. Renal function test, uric acid, and serum and urine electrolytes assay were performed. Second experiment: 18 male mice were used to evaluate the wound healing property of honey. Wounds were created on mice skin and treated daily with honey or Madecassol. Measurements of wounds were performed over a period of 12 days.

Results: The physical and chemical parameters of Tulkarm honey are within the limits of the European legislation and fulfilling the criteria described in the standard codex for honey. It contains antioxidant compounds and shows antioxidant activity. Oral honey increased creatinine clearance and urine volume, sodium, and chloride without causing hypokalemia or affecting blood urea, uric acid, or serum creatinine level. The diuretic activity of furosemide was associated with hypokalemia. Topical honey application enhanced wound closure when compared with the Madecassol application.

Conclusion: The study is the first to report that honey collected from Tulkarm has a considerable diuretic effect without affecting serum electrolytes or kidney function test and exhibits strong antioxidant activity and wound healing property.

Keywords: furosemide, honey, kidney, Madecassol, wounds.

An analysis of common foodborne parasitic zoonoses in slaughtered sheep and cattle in Tehran, Iran, during 2015-2018

Research (Published online: 24-10-2018)

20. An analysis of common foodborne parasitic zoonoses in slaughtered sheep and cattle in Tehran, Iran, during 2015-2018

Ali Pezeshki, Hadi Aminfar and Majid Aminzare

Veterinary World, 11(10): 1486-1490

ABSTRACT

Background and Aim: Cystic echinococcosis, Echinococcus granulosus, and liver flukes, such as Fasciola spp. and Dicrocoelium dendriticum, are important parasitic zoonoses, where they able to cause significant veterinary, medical, and economic problems. The present study was carried out to obtain the updated knowledge on the frequency of hydatidosis, fasciolosis, and dicrocoeliosis in the slaughtered sheep and cattle.

Materials and Methods: Information were collected from meat inspection records using systematically visual inspection, palpation, and incision of the visceral organs in the industrial abattoir in Tehran, the capital of Iran, between February 1, 2015, and January 31, 2018. For an analysis of the data, SPSS version 16 was applied.

Results: The hydatidosis infection in sheep and cattle was 2.48% and 2.25%, respectively. With respect to liver flukes, 0.62% and 0.25% sheep and cattle were infected by Fasciola spp., respectively; furthermore, 2.86% sheep and 0.79% cattle were positive for D. dendriticum.

Conclusion: The findings will provide considerable awareness for the future monitoring and control of these potentially important infections.

Keywords: dicrocoeliasis, fascioliasis, hydatidosis, sheep and cattle, slaughterhouse.

Nutritional potentialities of some tree leaves based on polyphenols and rumen in vitro gas production

Research (Published online: 23-10-2018)

19. Nutritional potentialities of some tree leaves based on polyphenols and rumen in vitro gas production

K. S. Giridhar, T. M. Prabhu, K. Chandrapal Singh, V. Nagabhushan, T. Thirumalesh, Y. B. Rajeshwari and B. C. Umashankar

Veterinary World, 11(10): 1479-1485

Aim: The study was conducted to evaluate eight tree leaves based on polyphenolic content and rumen in vitro incubation and gas production technique (RIVIGPT) for their nutritive potentiality.

Materials and Methods: Eight selected tree leaves, namely Sesbania grandiflora, Melia dubia, Dillenia spp., Artocarpus heterophyllus, Commiphora caudata, Moringa oleifera, Leucaena leucocephala, and Acacia auriculiformis, were selected for proximate composition, forage fiber fractions, total phenolics (TPs), non-tannin phenols (NTPs), total tannins (TTs), condensed tannins (CTs), and hydrolysable tannins (HTs); RIVIGP with and without polyethylene glycol (PEG); and in vitro dry matter digestibility (IVDMD) (modified in vitro two stage) analysis was conducted. On the basis of RIVIGPT, the in vitro digestible organic matter (IVDOM) and dry matter intake (DMI) was calculated.

Results: Crude protein (CP) content of tree leaves ranged from 9.59 to 25.81%, neutral detergent fiber (NDF) 28.16 to 53.33%, acid detergent fiber (ADF) 21.26 to 41.7%, acid detergent lignin (ADL) 3.62 to 21.98%, TP 1.83 to 17.35%, TT 0.40 to 15.47%, and CTs 0.02 to 15.26%. IVDMD (%) was ranged from 64.95 to 88.12. The mean metabolizable energy (ME) (MJ/Kg) of tree leaves estimated with and without PEG was 7.75±0.56 and 8.75±0.39, in vitro gas production at 24 h (IVGP24) (ml) 31.06±4.14 and 37.09±2.64, initial gas production (a) (ml) 0.49±0.63 and 1.33±0.72, potential gas production (D) (ml) 38.74±4.27 and 43.79±2.44, rate of gas production (k) (h-1) 0.11±0.02 and 0.11±0.013, t1/2 (ml) 9.81±2.41 and 7.42±0.80, in vitro gas production at 96 h IVGP96 (ml) 39.50±4.430 and 45.14±2.65, the predicted IVDOM (%) 55.44±4.15 and 61.98±3.03, and DMI (g/Kg W0.75) 103.1±14.76 and 104.3±10.16, respectively. The addition of PEG showed an improvement in IVGP24, IVGP96, ME, predicted IVDOM, and predicted DMI. CP was positively correlated with ME, IVGP24, IVGP96, a+b, k (r=0.749, p<0.05), IVDMD, IVDOM, and DMI (r=0.838, p<0.05) and negatively correlated with a and t1/2. NDF, ADF, and ADL contents were negatively correlated with ME (r=0.899, p<0.05), IVGP24 (r=-0.867, p<0.05), IVGP96 (r=-0.858, p<0.05), a+b (p<0.05), k (r=-0.828, p<0.05), IVDMD, IVDOM (r=-0.853, p<0.05), and DMI and positively correlated with a and t1/2. TP, TT, and CT were negatively correlated with ME, IVGP, IVGP96, a+b, k, IVDMD, IVDOM, and DMI and positively correlated with a (r=0.808, p<0.05) and t1/2. ME (MJ/Kg) was positively correlated with IVGP24 (r=0.938, p<0.05), IVGP96 (r=0.875, p<0.05), a+b (r=0.813, p<0.05), k (r=0.731, p<0.05), IVDMD, IVDOM (r=0.985, p<0.05), and DMI (r=0.727, p<0.05) and negatively correlated with a and t1/2.

Conclusion: In the present study, the potentiality of tree leaves was assessed based on CP, ADF, ADL, TP, CT, IVGP, ME, IVDMD, predicted IVDOM, and predicted DMI. Based on this, it can be concluded that S. grandiflora, M. dubia, M. Oleifera, and L. leucocephala were graded as best; A. heterophyllus and C. caudata as moderate; and Dillenia spp. and A. auriculiformis as lowest potential ruminant feed.

Keywords: chemical composition, in vitro, in vitro dry matter digestibility, in vitro digestible organic matter, metabolizable energy, polyethylene glycol, rumen in vitro incubation and gas production, ruminants, tannins, tree leaves.

Structural uterine changes in postpartum endometritis in cows

Research (Published online: 22-10-2018)

18. Structural uterine changes in postpartum endometritis in cows

S. M. Suleymanov, B. V. Usha, Yu. A. Vatnikov, E. D. Sotnikova, Eu. V. Kulikov, V. I. Parshina, M. V. Bolshakova, M. U. Lyshko and E. V. Romanova

Veterinary World, 11(10): 1473-1478

ABSTRACT

Aim: The purpose of this work was to study the dynamics of structural manifestations of acute cases of postpartum endometritis in cows.

Materials and Methods: The light and electron microscopy methods were used when studying structural changes in the endometrium in case of postpartum endometritis in seven cows. Sections of endometrial specimens for light microscopy, 5-7 μm thick, were stained with hematoxylin and eosin and also by Van Gieson's. For electron microscopy, semi-thin sections were stained with Azur-2 in combination with basic fuchsin, as well as contrasting by lead citrate and uranyl acetate.

Results: As a result of the study, we have established the following: Necrobiosis of the epithelial layer of the mucosa, cellular infiltration with shaped elements of blood in the functional layer, swelling of the cells of the uterine gland, and single microbial cells on the surface of the mucosa. We have noted edema of the stroma of the functional layer of the endometrium, swelling of the epithelial layer of the endometrial mucosa, and swelling of fibroblastic and lymphoid cells. Ultrastructural changes in endometrial cells in case of acute postpartum endometritis in cows are accompanied by the destruction of microvilli on the apical surface of the epithelium, an abundance of coccal microflora on the surface of the epithelium, necrobiosis of epithelial cells, and partial edema of the nucleus, and cytoplasm of the histiocyte.

Conclusion: We had established that acute purulent-catarrhal dystrophic processes were observed in the structural organization of the endometrium. In the depth of catarrhal mucus on the surface of the endometrium, there was an abundance of bacterial flora, with diplococci being prevalent. In ultrastructural organization of the endometrium, we observed deep dystrophic and necrobiotic processes in the parenchyma and endometrial stroma, as well as exudative processes with a change in the integrity of the microcirculatory bed. Thus, to prevent an inflammatory process from turning into a latent form, it is necessary to detect acute postpartum endometritis promptly using diagnostic methods taking into account the obtained parameters of the dynamics of structural changes in the uterine tissues.

Keywords: cows, histological, macroscopic, postpartum endometritis, subclinical endometritis, ultrastructural changes of the endometrium.

Opportunities and challenges associated with fecal progesterone metabolite analysis

Review (Published online: 20-10-2018)

17. Opportunities and challenges associated with fecal progesterone metabolite analysis

Innocent Damudu Peter, Abd Wahid Haron, Faez Firdaus Abdullah Jesse, Mokrish Ajat, Mark Hiew Wen Han, Wan Nor Fitri, Muhammad Sanusi Yahaya and Mohammed Saad M. Alamaary

Veterinary World, 11(10): 1466-1472

ABSTRACT

Conventionally, plasma or milk progesterone evaluations are used to determine the reproductive status of female animals. Collection of such samples is often associated with difficulties of animal handling and restraint. Measurable quantities of progesterone metabolites are found in feces of animals. Their concentrations are known to be well correlated to plasma progesterone levels and are, therefore, used as non-invasive samples for assessing reproductive function in a wide range of animal species. Although the analysis of fecal progesterone metabolites has been widely accepted in many laboratories, several factors are known to affect the results from this valuable analytical technique. Some of these factors include storage/ transportation media for fecal samples, type of solvent that is used for extraction of progesterone metabolites from feces, and the type and sensitivity of an assaying technique employed. Although fecal progesterone metabolites analysis is associated with some difficulties, it can effectively be used to monitor reproductive function in a wide range of animal species. This review aims to highlight the usefulness of fecal progesterone metabolite analysis as a non-invasive technique in monitoring reproductive function in animals. The article mainly focuses on the many opportunities and challenges associated with this analytical technique.

Keywords: non-invasive methods, progesterone metabolite, progesterone, reproductive cycles.

Antibiotic Susceptibility profile of Staphylococcus aureus isolated from sausages in Meknes, Morocco

Research (Published online: 19-10-2018)

16. Antibiotic Susceptibility profile of Staphylococcus aureus isolated from sausages in Meknes, Morocco

Abdelaziz Ed-Dra, Fouzia Rhazi Filali, Aziz Bouymajane, Faouzia Benhallam, Abdellah El Allaoui, Abdellah Chaiba and Filippo Giarratana

Veterinary World, 11(10): 1459-1465

ABSTRACT

Background and Aim: Staphylococcus aureus is one of the most common causes of foodborne disease worldwide, due to the consumption of food contaminated by their toxins. This study aimed to determine the prevalence and the antimicrobial resistance of S. aureus isolated from sausages in Meknes city of Morocco.

Materials and Methods: A total of 156 samples (Beef sausages, Turkey sausages, and Artisanal sausages "Merguez") were collected from different shopping sites (butchery, supermarket, street vendors, and weekly market "Souk") and used for the isolation of S. aureus. All the isolated strains were tested for their antimicrobials resistance to 16 antibiotics.

Results: Our results showed the presence of S. aureus in 63 samples (40.38%). Furthermore, the antimicrobial resistance study showed that 84.13% of isolated S. aureus were resistant to streptomycin, 76.20% to tetracycline, 42.86% to ampicillin, 41.27% to doxycycline, 38.1% to penicillin G, and 19.05% to chloramphenicol with the presence of 25 different phenotypic profiles. However, all isolated strains were sensitive to oxacillin, cefoxitin, gentamicin, and vancomycin.

Conclusion: The findings of this study revealed consumption of sausages as a potential risk of foodborne poisonings because of its contamination with the multi-resistant strains of S. aureus. Moreover, this contamination is related to the season, sampling sites and the origin of the raw material.

Keywords: antimicrobial resistance, foodborne disease, infection, sausages, Staphylococcus aureus.

Occurrence of Escherichia coli carrying Shiga toxin-producing genes in buffaloes on smallholdings in Bangladesh

Research (Published online: 19-10-2018)

15. Occurrence of Escherichia coli carrying Shiga toxin-producing genes in buffaloes on smallholdings in Bangladesh

Mukta Das Gupta, Arup Sen and Ashutosh Das

Veterinary World, 11(10): 1454-1458

ABSTRACT

Background and Aim: Shiga toxin-producing Escherichia coli (STEC) has emerged as significant foodborne pathogens. Ruminants are the primary reservoir of the zoonotic STEC. In Bangladesh, previous studies reported the presence of STEC in cattle, goat, and sheep; however, there is little information about STEC carriage by buffaloes. This study aimed to determine the occurrence of STEC in healthy (absence of clinical signs and symptoms) buffaloes on smallholdings in Bangladesh and to assess the antimicrobial resistance pattern of identified STEC isolates.

Materials and Methods: A total of 100 rectal swab samples were obtained from randomly selected buffaloes on 40 smallholdings in Chittagong Division, Bangladesh. Samples were subjected to bacteriological screening to identify E. coli. All E. coli isolates were examined for the presence of the Shiga toxin-producing genes - Shiga toxin 1 (stx1) and Shiga toxin 2 (stx2) using polymerase chain reaction. The antimicrobial susceptibility of identified STEC isolates was tested using the disk diffusion method.

Results: Results show that 71 fecal samples were positive for E. coli in bacteriological screening. The proportion of buffaloes harboring STEC isolates was 11% (11/100) (95% confidence interval [CI] 6.1-18.8], of which 7% (7/100) (95% CI 3.2- 13.9) and 4% (4/100) (95% CI 1.2-10.2) carried stx1 and stx2 genes, respectively. Antibiogram revealed that 91% (10/11), 73% (8/11), 55% (6/11), and 55% (6/11) STEC isolates were resistant to tetracycline, sulfamethoxazole-trimethoprim, erythromycin, and ampicillin, respectively. In contrast, 91% (10/11) STEC isolates were sensitive to ciprofloxacin, chloramphenicol, and gentamicin, whereas 73% (8/11) isolates were sensitive to ceftriaxone.

Conclusion: This study highlights, for the first time, a significant proportion of fecal samples from healthy buffaloes on smallholdings in Bangladesh harboring antimicrobial-resistant STEC. Transmission of antimicrobial-resistant STEC from buffaloes to humans could pose an added risk to public health in rural Bangladesh.

Keywords: antimicrobial resistance, buffalo, Shiga toxin-producing Escherichia coli.

Antibacterial effect of ozonated water against methicillin-resistant Staphylococcus aureuscontaminating chicken meat in Wasit Province, Iraq

Research (Published online: 18-10-2018)

14. Antibacterial effect of ozonated water against methicillin-resistant Staphylococcus aureuscontaminating chicken meat in Wasit Province, Iraq

Manal H. G. Kanaan

Veterinary World, 11(10): 1445-1453

ABSTRACT

Background and Aim: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most recognized "superbugs" and a common cause of community-associated and nosocomial infections; furthermore, when chicken meat is considered a good growth medium for S. aureus to make a plausible vehicle to propagate MRSA, then this study was conducted to evaluate the efficiency of ozonated water (0.5 ppm) in the elimination or reduction of MRSA contaminating fresh and frozen chicken meat sold in local markets in the Wasit Province.

Materials and Methods: A total of 72 samples of fresh and frozen chicken meat were randomly collected from dissimilar native markets: Fresh chicken meat (n=32) and frozen chicken meat (n=40). Isolation and identification of MRSA isolates were conducted using standard bacteriological, biochemical, RapID™ Staph Plus System (Remel, R8311009), and latex agglutination tests such as Dry SPOT Staphytect Plus (Oxoid, DR0100M) and PBP2' Test Kit (Oxoid, DR0900A). The generation of ozone (O3) was carried out using O3 generator (A2Z/AQUA-6, USA), and its concentration (ppm) in water was determined using CHE-Mets®-Kit, USA.

Results: A total of 39 (54.2%) of 72 fresh and frozen chicken meat were positive for S. aureus; of those 39 positive samples, 13 (33.3%) were identified as MRSA. The antibiotic sensitivity test results revealed that all MRSA isolates had multiple resistance to at least four antimicrobial agents for which these isolates had 12 antibiotic resistance patterns. Results of O3 treatment in MRSA isolate contaminating 13 of both fresh and frozen chicken meat samples showed that, after treatment with ozonated water (0.5 ppm/4°C), the overall negative samples were 23.1% and 69.2% for 30 and 45 min, respectively. The decrease in the percentage of positive samples was very significant from a public health perspective. Furthermore, the antimicrobial efficacy of ozonated water (0.5 ppm) on the reduction of the MRSA count (log10 colony-forming units [CFU]/ml) was assessed in four positive samples of fresh and frozen chicken meat, and the results revealed that, after treatments, the overall reduction was 2-4 log10 (CFU/ml) after 45 min. This reduction is highly significant from a public health perspective.

Conclusion: From the data obtained from this study, it can be concluded that fresh and frozen chicken meat sold in the different markets of Wasit Province was highly contaminated by S. aureus during the study period with a total prevalence of 54.2%; among those, 33.3% were recognized as MRSA. Under the conditions described in the present study, O3 at the concentration of 0.5 ppm is highly effective in reducing the number of MRSA-positive samples and the number decreased with increased exposure time to ozonated water at the same concentration. These findings indicated that O3 treatment might constitute the basis for an alternative method to reduce meat contamination with foodborne pathogens such as MRSA.

Keywords: antibacterial effect, chicken meat, methicillin-resistant Staphylococcus aureus, ozonated water, Wasit Province.

Antagonistic effect of ursolic acid on Staphylococcal biofilms

Research (Published online: 17-10-2018)

13. Antagonistic effect of ursolic acid on Staphylococcal biofilms

J. Shiva Jyothi, Kalyani Putty, Y. Narasimha Reddy, K. Dhanalakshmi and M. A. Hannan Umair

Veterinary World, 11(10): 1440-1444

ABSTRACT

Aim: The present study was carried out to study the effect of ursolic acid (UA) as a potential anti-biofilm agent in dispersing the biofilm generated by Staphylococcus aureus isolated from milk samples of crossbred dairy cows on the day of drying. Further, in the S. aureus isolates, the presence of intracellular adherence gene locus involved in biofilm production (icaD) was investigated.

Materials and Methods: A total of 50 S. aureus strains were isolated over a period of 3 months from 200 milk samples collected from crossbred dairy cows on the day of drying. These isolates were subjected for biofilm detection by Congo red agar (CRA), microtiter plate assay (MTP), and polymerase chain reaction specific for icaD gene. The antagonistic effect of biofilm formation by UA was studied using different concentrations (30 μg/ml and 60 μg/ml) of UA and compared with the control group.

Results: Among the 50 S. aureus subjected for biofilm detection, 34 and 40 isolates were detected as biofilm agents by CRA and MTP methods, respectively. The in vitro studies on the effect of UA in inhibiting biofilm formation by S. aureus using MTP assay showed 71.5% and 48.6% inhibition at UA concentrations of 60 μg/ml and 30 μg/ml, respectively, with a significant difference (p<0.05) between the treated and untreated isolates, which was further evident by scanning electron microscopy. Interestingly, the isolates that were tested to be resistant through Antibiotic Sensitivity Test to commonly used antibiotics were found to be sensitive to all the tested antibiotics following UA treatment at both the tested concentrations. Furthermore, molecular detection of icaD gene for biofilm detection revealed that all the isolates that were positive by MTP had icaD gene.

Conclusion: Increased incidence of biofilm agents in dairy infections must be considered as an alarming situation. UA treatment significantly enhanced the sensitivity of the microbial pathogens to commonly used antibiotics. Hence, attention must be paid toward implementation of new strategies such as therapeutic regimes with a combination of antibiotic and anti-biofilm agents for effective treatment of infections in dairy farms.

Keywords: biofilms, icaD, Staphylococcus aureus, ursolic acid.

Supplementation of whole grain flaxseeds (Linum usitatissimum) along with high cholesterol diet and its effect on hyperlipidemia and initiated atherosclerosis in Wistar albino male rats

Research (Published online: 17-10-2018)

12. Supplementation of whole grain flaxseeds (Linum usitatissimum) along with high cholesterol diet and its effect on hyperlipidemia and initiated atherosclerosis in Wistar albino male rats

H. Srinivasa Naik, Ch. Srilatha, K. Sujatha, B. Sreedevi and T. N. V. K. V. Prasad

Veterinary World, 11(10): 1433-1439

ABSTRACT

Background and Aim: Flaxseeds are known to have varying antihypercholesterolemic and antiatherogenic activity due to its lignan secoisolariciresinol diglucoside, alpha-linolenic acid, and omega-3 fatty acids. The beneficial effect of whole grain dietary flaxseed was evaluated experimentally in high cholesterol diet (HCD)-fed Wistar albino rats.

Materials and Methods: Male Wistar albino rats (200 g) were divided into four groups of 12 rats each. Group I rats kept as control and given basal rat chew diet, Group II as positive control for induction of hypercholesterolemia and atherosclerosis by addition of 1% cholesterol and 15% saturated edible oil to the 1000 g of standard rat chew diet (HCD), Group III rats fed with whole grain flaxseed powder at 7.5 g/kg of rat/day in the standard rat chew diet and kept as flaxseed control, and Group IV rats supplemented with flaxseed at 7.5 g/kg of rat/day along with HCD and maintained for 90 days.

Results: Group II rats revealed significantly (p<0.05) higher total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and very LDL-C and significantly (p<0.05) reduced levels of high-density lipoprotein cholesterol (HDL-C), whereas tissue antioxidants such as catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S transferase (GST) were significantly (p<0.05) reduced, and lipid peroxidation products of thiobarbituric acid reactive substances (TBARS) level were nonsignificantly (p<0.05) increased in the heart and liver tissues. Flaxseeds supplementation along with HCD significantly ameliorated the serum levels of TC, TG, LDL-C, and HDL-C along with cellular antioxidant enzymes such as catalase, SOD, GPx, GR, GST, and non-significant amelioration of TBARS in the heart and liver tissues compared to Group II rats. Majority of the histopathologically initiated atherosclerotic changes in the aorta and fatty change in the liver of Group II were not observed in the flaxseed supplemented Group IV; however, interestingly proliferation of endothelial cells with new vascular channel formation in the liver and in between cardiac muscle fibers was observed in Group I and Group IV rats.

Conclusion: The present study established the hypercholesterolemia with initiated atherosclerotic lesion in the aorta but unable to establish the atheromatous plaque in the aorta. Flaxseed supplementation along with HCD showed significant antihypercholesterolemic effect and ameliorated the changes of initiated atherosclerosis in the aorta. It needs further studies to explore all the possible beneficial effects and angiogenic properties of flaxseeds in the laboratory animals and human trials.

Keywords: atherosclerosis, flaxseeds, hepatic steatosis, hyperlipidemia, tissue antioxidants.

In vitro antibacterial effects of zinc oxide nanoparticles on multiple drug-resistant strains of Staphylococcus aureus and Escherichia coli: An alternative approach for antibacterial therapy of mastitis in sheep

Research (Published online: 16-10-2018)

11. In vitro antibacterial effects of zinc oxide nanoparticles on multiple drug-resistant strains of Staphylococcus aureus and Escherichia coli: An alternative approach for antibacterial therapy of mastitis in sheep

Myassar Alekish, Zuhair Bani Ismail, Borhan Albiss and Sara Nawasrah

Veterinary World, 11(10): 1428-1432

ABSTRACT

Aim: The aim of the study was to evaluate the antibacterial effects of zinc oxide nanoparticles (ZnO-NPs) and its possible alternative use for the treatment for mastitis in sheep and to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ZnO-NPs against multidrug-resistant Staphylococcus aureus and Escherichia coli strains isolated from subclinical mastitis cases in sheep.

Materials and Methods: A total of 50 pooled milk samples were collected from ewes with subclinical mastitis. Milk samples were cultured using standard laboratory techniques, and multidrug-resistant bacterial strains were determined using the Kirby-Bauer disk diffusion method. The MIC and MBC of ZnO-NPs were determined against isolated multidrug-resistant S. aureus and E. coli strains using microwell dilution method.

Results: A total of 43 different bacterial isolates were recovered from milk samples of ewes affected with subclinical mastitis. Isolated strains of S. aureus and E. coli were found resistant to three or more common antibacterial agents and were used to determine the MIC and MBC of ZnO-NPs. The MIC and MBC values of ZnO-NPs were significantly lower for S. aureus than that for E. coli. The MIC and MBC of ZnO-NPs against S. aureus were 3.9 μg/ml and 7.81 μg/ml, respectively, while for E. coli, the MIC and MBC of ZnO-NPs were 31.25 μg/ml and 62.5 μg/ml, respectively.

Conclusion: Results of this study indicate the potential antibacterial effects of ZnO-NPs against multidrug-resistant S. aureus and E. coli isolated from ovine subclinical mastitis at concentrations of 3.9 μg/ml and 31.25 μg/ml, respectively.

Keywords: alternative therapy, antibiotics, mastitis, nanotechnology, sheep, zinc oxide.

Detection, characterization, and antibiogram of extended-spectrum beta-lactamase Escherichia coliisolated from bovine milk samples in West Bengal, India

Research (Published online: 16-10-2018)

10. Detection, characterization, and antibiogram of extended-spectrum beta-lactamase Escherichia coliisolated from bovine milk samples in West Bengal, India

Kunal Batabyal, Abhiroop Banerjee, Susmita Pal, Samir Dey, Siddhartha Narayan Joardar, Indranil Samanta, Devi Prasad Isore and Abhishek Dharm Singh

Veterinary World, 11(10): 1423-1427

ABSTRACT

Background: Milk is considered as complete food and an important part of human diet throughout the world including India. Bacterial contamination of milk such as Escherichia coli due to unhygienic condition and poor udder health can cause infections, especially in infants and elders or in immunocompromised persons. Possession of antimicrobial resistance genes by commensal bacteria present in milk makes the issue more serious.

Aim: The study was aimed to isolate and characterize extended-spectrum beta-lactamase (ESBL)-producing E. coli from milk samples collected from different parts of West Bengal, India, to assess the potential risk associated with the food.

Materials and Methods: Around 182 milk samples were collected from apparently healthy cows reared by organized dairy farms in West Bengal. E. coli was isolated from collected samples as per standard methods followed by serotyping. The detection of ESBL-producing E. coli was done both phenotypically and genotypically by detecting the presence of blaCTX-M gene. Antibiogram of the ESBL-positive isolates was done using common 12 antibiotics by disc diffusion method.

Results: A total of 22 (12.1%) samples were found to be positive for E. coli in this study. Different serotypes such as O11, O20, O22, O34, O35, O128, O149, and UT were isolated from the collected samples. 12 (54.5%) E. coli strains showed the capability of producing ESBL, both phenotypically and genotypically with the presence of blaCTX-M gene. Antibiogram of these ESBL-positive isolates revealed the drugs such as colistin (100%), levofloxacin (83.33%), and imipenem (66.67%) to be highly sensitive against this pathogen but drugs such as cefotaxime (100%), ceftazidime (91.67%), amoxicillin/clavulanic acid (83.33%), tetracycline (75.00%), and gentamicin (58.33%) to be very much resistant.

Conclusion: More than 50% of the E. coli strains prevalent in the bovine milk samples were positive for ESBL production and are resistant to most of the common antimicrobials which may be alarming for human health.

Keywords: antibiogram, blaCTX-M, bovine milk, extended-spectrum beta-lactamase, Escherichia coli.