γ-Aminobutyric acid production by selected lactic acid bacteria isolate of an Indonesian indigenous fermented buffalo milk (dadih) origin

Research (Published online: 30-08-2019)

26. γ-Aminobutyric acid production by selected lactic acid bacteria isolate of an Indonesian indigenous fermented buffalo milk (dadih) origin

Harnentis Harnentis, Nurmiati Nurmiati, Yetti Marlida, Frederick Adzitey and Nurul Huda

Veterinary World, 12(8): 1352-1357

ABSTRACT

Aim: This study aimed at optimizing γ-aminobutyric acid (GABA) production using lactic acid bacteria (LAB) of an Indonesian indigenous fermented buffalo milk (dadih) origin. This study utilized LAB previously cultured from dadih that has the ability to produce GABA.

Materials and Methods: The study started with the identification of selected LAB by 16S rRNA, followed by optimization of GABA production by culture conditions using different initial pH, temperature, glutamate concentration, incubation time, carbon, and nitrogen sources. 16S rRNA polymerase chain reaction and analysis by phylogenetic were used to identify Lactobacillus plantarum (coded as N5) responsible for the production of GABA.

Results: GABA production by high-performance liquid chromatography was highest at pH of 5.5, temperature of 36°C, glutamate concentration of 500 mM, and incubation time of 84 h. Peptone and glucose served as the nitrogen and carbon sources, respectively, whereas GABA was produced at optimum fermentation condition of 211.169 mM.

Conclusion: Production of GABA by L. plantarum N5 was influenced by initial pH of 5.5, glutamic acid concentration, nitrogen source, glucose as carbon source, and incubation temperature and time.

Keywords: fermented buffalo milk, Indonesian indigenous product, lactic acid bacteria, γ-aminobutyric acid.

Coleus amboinicus extract increases transforming growth factor-1β expression in Wistar rats with cisplatin-induced nephropathy

Research (Published online: 28-08-2019)

25. Coleus amboinicus extract increases transforming growth factor-1β expression in Wistar rats with cisplatin-induced nephropathy

Iwan Sahrial and Rondius Solfaine

Veterinary World, 12(8): 1346-1351

ABSTRACT

Background and Aim: Coleus amboinicus (CA) plants are known to exert antibacterial and anti-inflammatory effects and demonstrate antiproliferative effects against cancer cells. This study aimed to investigate the activity of CA extract on the expression of transforming growth factor-1β (TGF-1β) in cisplatin-induced nephropathy in Wistar rats (Rattus norvegicus).

Materials and Methods: CA was obtained from fresh leaves of CA and was extracted using 96% ethanol maceration. This blinded, controlled, randomized post-test study assigned 24 Wistar rats to three groups (n=8). Negative controls received normal saline (P0), nephropathy was induced in rats by cisplatin (5 mg/kg, IP) (P1), and treated with ethanolic coleus extract (500 mg/kg, PO) (P2), respectively, for 7 days. Nephropathy was induced on the 4th day. All rats were sacrificed on the 8th day for blood and kidney sample collection. Concentrations of blood urea nitrogen (BUN), creatinine, and alkaline phosphatase were analyzed using colorimetric analysis. A semi-quantitative analysis was performed on sectioned kidneys to determine the numbers of positive cells for TGF-1β expression and to evaluate structural and functional alterations in the kidneys using histopathological and immunohistochemical staining.

Results: The concentrations of BUN, creatinine, and alkaline phosphatase from blood samples in the treatment group were significantly lower than those of the control group. Morphological evaluation of the tubular interstitium and glomeruli revealed that necrotic, degenerating, and infiltration of cells significantly decreased in the treatment group compared to the control group. The mean immunostaining scores indicating the presence of TGF-1β were 7.8 in the ethanolic coleus extract group, 3 in the induction group, and 2.3 in the control group. The expression scores for TGF-β1 were significantly different between the ethanolic coleus extract treatment and control group.

Conclusion: Our results suggest that in Wistar rats with cisplatin-induced nephropathy, CA extract inhibits pathological lesions by regulating the renal expression of TGF-1β in areas containing the renal tubules and glomeruli. 

Keywords: cisplatin, Coleus amboinicus, nephropathy, transforming growth factor-1β.

Morphological and molecular identification of Pfenderius heterocaeca (Trematode: Paramphistomoidea) from Sumatran elephant (Elephas maximus sumatranus)

Research (Published online: 28-08-2019)

24. Morphological and molecular identification of Pfenderius heterocaeca (Trematode: Paramphistomoidea) from Sumatran elephant (Elephas maximus sumatranus)

Lintang Winantya Firdausy, Rahmania Prahardani, Lu'lu' Sahara Wusahaningtyas, Soedarmanto Indarjulianto, Muhammad Wahyu, Muhammad Tauhid Nursalim and Wisnu Nurcahyo

Veterinary World, 12(8): 1341-1345

ABSTRACT

Background and Aim: Paramphistomiasis is common in tropical countries such as Indonesia and affects livestock and various endemic wild animals such as Sumatran elephants. However, the specific species of paramphistomoid worm that causes paramphistomiasis are rarely reported. The study aims at identifying paramphistomoid worm that infects Sumatran elephants.

Materials and Methods: Flukes were collected from the feces of five semi-captive Sumatran elephants that lived at Tegal Yoso Elephant Response Unit in Way Kambas National Park, in 2018, after treatment of oxyclozanide 1 g at the dose of approximately 5-8 mg/kg of body weight. Eight paramphistomoid worms were flattened and stained in Semichon's carmine for morphological identification, and five other worms were used for molecular identification at second internal transcribed spacer (ITS-2) of ribosomal deoxyribonucleic acid sequence.

Results: Forty-five flukes were collected from five Sumatran elephants in Lampung, Indonesia. Eight paramphistomoid worms were morphologically identified as Pfenderius heterocaeca> and five isolates did not show any variation in ITS-2. Phylogenetic analysis showed that there was a close genetic relationship between our sample and Chiorchis fabaceus that had a family similar to the samples.

Conclusion: Based on the morphological and molecular characteristics, the paramphistomoids found in Sumatran elephant on Way Kambas National Park are P. heterocaeca.

Keywords: internal transcribed spacer-2, paramphistomiasis, Pfenderius spp., Sumatran elephant.

Immune cell pathology in rabbit hemorrhagic disease

Research (Published online: 28-08-2019)

23. Immune cell pathology in rabbit hemorrhagic disease

Anna Babken Semerjyan, Mariam Armenak Sargsyan, Hranush Harutyun Arzumanyan, Lina Hayrapet Hakobyan, Liana Onik Abroyan, Zara Babken Semerjyan, Aida Sergey Avetisyan, Elena Michael Karalova, Davit Mihran Manukyan, Hripsime Shavarsh Matevosyan, Nikolay Fyodor Krasnikov and Zaven Alexandr Karalyan

Veterinary World, 12(8): 1332-1340

ABSTRACT

Aim: The aim of this research was to study the effect of rabbit hemorrhagic disease virus (RHDV) on the host immune response by examining the cellular composition/pathology of lymphoid organs and serum levels of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ).

Materials and Methods: Nine adult rabbits were inoculated with 1 ml of 10% infected liver homogenate, and three rabbits served as controls. The rabbit hemorrhagic disease (RHD)-induced animals were studied on 3 consecutive days post-infection. Diagnosis of RHD was made through routine hemagglutination tests and the polymerase chain reaction. Blood smears and tissue samples from bone marrow (BM), spleen, lymph nodes, and liver were analyzed for cell composition and cytopathology. Serum levels of TNF-α and IFN-γ were measured by enzyme-linked immunosorbent assay.

Results: RHD showed a decreased absolute cell count of blood as well as lymph nodes, spleen, and BM cell populations with marked left shift. This was seen as a progressive rise in immature and blast cells. Quantitative cellular changes were accompanied by an increase in specific inflammatory cytokines. Immunocytopathological alterations were evidenced by: Vacuolized, hyperactivated tissue macrophages, finding of Dohle bodies in neutrophils, and activated lymphocytes with increased nuclear-cytoplasmic ratio. Cytoplasmic eosinophilic viral inclusions found in tissue (liver, spleen, and BM) macrophages were shown for the 1st time in RHD. Megakaryocytic emperipolesis was a common feature of RHD.

Conclusion: These studies suggest that RHDV induces pathology in leukocytes due to hyperactivation with left shift (toward immature stages of the different cell lineages). Macrophages are increased in number and show an expressed cytopathic effect often accompanied by viral eosinophilic cytoplasmic inclusions. They also developed a secretory activation (increased levels of pro-inflammatory cytokines).

Keywords: cytopathology, emperipolesis, eosinophilic viral inclusions, immune response, macrophages, rabbit hemorrhagic disease virus.

Non-genetic factors affecting pre-weaning growth and morphometric traits in Assam Hill goat

Research (Published online: 25-08-2019)

22. Non-genetic factors affecting pre-weaning growth and morphometric traits in Assam Hill goat

L. Sarma, N. Nahardeka, R. N. Goswami, A. Aziz, G. Zaman, A. Das and F. Akhtar

Veterinary World, 12(8): 1327-1331

ABSTRACT

Aim: This study aimed to determine the genetic and non-genetic factors affecting pre-weaning body weight (BW) and morphometry in Assam Hill goat along with the genetic parameters.

Materials and Methods: The detailed information in respect of BW and body measurements of 960 animals at birth and 3 months of age belonging to three different populations of Assam Hill goat maintained at field units, namely, Batabari, Nahira, and Tetelia under "All India Coordinated Research Project on Goat Improvement" were utilized in the present study. The data were analyzed using least squares technique.

Results: The least squares means for BW, height at withers (HW), heart girth (HG), and body length (BL) were 1.166±0.008 kg, 26.198±0.070 cm, 26.695±0.096 cm, and 29.482±0.119 cm at birth and 4.590±0.083 kg, 36.850±0.105 cm, 40.741±0.115 cm, and 39.703±0.108 cm at 3 months of age, respectively. Location had a significant effect on BW, HW, and BL at both birth and 3 months and on HG at 3 months of age. Season of birth exerted significant effect only on BL at birth, whereas the significant effect of sex was observed on HG and BL at 3 months of age. The heritability estimates for BW and body measurements were moderate indicating the scope of selection. The phenotypic and genetic correlations among BWs and body measurements at birth and 3 months of age were positive in direction and high in magnitude.

Conclusion: On the basis of the present findings, it could be concluded that the weaning weight of kids can be considered for the selection of parent stock to increase productivity and eventually the economic efficiency. Further, animals with higher body measurements at initial phases of growth will perform better with respect to even BW at later stages of growth.

Keywords: Assam Hill goat, genetic parameters, morphometric traits, non-genetic factors, pre-weaning growth.

In vitro study of ivermectin efficiency against the cattle tick, Rhipicephalus (Boophilus) annulatus, among cattle herds in El-Beheira, Egypt

Research (Published online: 25-08-2019)

21. In vitro study of ivermectin efficiency against the cattle tick, Rhipicephalus (Boophilus) annulatus, among cattle herds in El-Beheira, Egypt

Gaber E. Batiha, Ali H. El-Far, Amany A. El-Mleeh, Abdelwahab A. Alsenosy, Eman K. Abdelsamei, Mohamed M. Abdel-Daim, Yasser S. El-Sayed and Hazem M. Shaheen

Veterinary World, 12(8): 1319-1326

ABSTRACT

Background and Aim: Ivermectin (IVM) has been used in veterinary practice to control different parasitic infestations over the past two decades. This study aimed to re-assess the acaricidal effects of IVM, as well as to evaluate its efficacy against Rhipicephalus (Boophilus) annulatus by determining the mortality rate, γ-aminobutyric acid (GABA) level, and oxidative/ antioxidative homeostasis (malondialdehyde [MDA] levels and glutathione S-transferase [GST] activities).

Materials and Methods: Adult females of Rhipicephalus (Boophilus) annulatus were picked from cattle farms in El-Beheira Governorate, Egypt. Ticks were equally allocated to seven experimental groups to assess the acaricidal potential of IVM chemotherapeutics in controlling R. (B.) annulatus. IVM was prepared at three concentrations (11.43, 17.14, and 34.28 μM of IVM).

Results: Mortality rate was calculated among the treated ticks. In addition, GABA, GST, and MDA biomarker levels were monitored. The data revealed a noticeable change in GST activity, a detoxification enzyme found in R. (B.) annulatus, through a critical elevation in mortality percentage.

Conclusion: IVM-induced potent acaricidal effects against R. (B.) annulatus by repressing GST activity for the initial 24 h after treatment. Collectively, this paper reports the efficacy of IVM in a field population of R. (B.) annulatus in Egypt.

Keywords: acaricidal, glutathione S-transferase, ivermectin, malondialdehyde, Rhipicephalus (Boophilus) annulatus, γ-aminobutyric acid.

Molecular epidemiology and antimicrobial resistance of Salmonella isolates from broilers and pigs in Thailand

Research (Published online: 23-08-2019)

20. Molecular epidemiology and antimicrobial resistance of Salmonella isolates from broilers and pigs in Thailand

Dusadee Phongaran, Seri Khang-Air and Sunpetch Angkititrakul

Veterinary World, 12(8): 1311-1318

ABSTRACT

Aim: This study aimed to determine the prevalence and antimicrobial resistance pattern of Salmonella spp., and the genetic relatedness between isolates from broilers and pigs at slaughterhouses in Thailand.

Materials and Methods: Fecal samples (604 broilers and 562 pigs) were collected from slaughterhouses from April to July 2018. Salmonella spp. were isolated and identified according to the ISO 6579:2002. Salmonella-positive isolates were identified using serotyping and challenged with nine antimicrobial agents: Amoxicillin/clavulanate (AMC, 30 μg), ampicillin (AMP, 10 μg), ceftazidime (30 μg), chloramphenicol (30 μg), ciprofloxacin (CIP, 5 μg), nalidixic acid (NAL, 30 μg), norfloxacin (10 μg), trimethoprim/sulfamethoxazole (SXT, 25 μg), and tetracycline (TET, 30 μg). Isolates of the predominant serovar Salmonella Typhimurium were examined for genetic relatedness using pulsed-field gel electrophoresis (PFGE).

Results: Salmonella was detected in 18.05% of broiler isolates and 37.54% of pig isolates. The most common serovars were Kentucky, Give, and Typhimurium in broilers and Rissen, Typhimurium, and Weltevreden in pigs. Among broilers, isolates were most commonly resistant to antibiotics, NAL, AMP, TET, AMC, and CIP. Pig isolates most commonly exhibited antimicrobial resistance against AMP, TET, and SXT. Based on PFGE results among 52 S. Typhimurium isolates from broilers and pigs, a high genetic relatedness between broiler and pig isolates (85% similarity) in Cluster A and C from PFGE result was identified.

Conclusion: The results revealed high cross-contamination between these two animal species across various provinces in Thailand.

Keywords: antimicrobial resistance, broilers, pigs, pulsed-field gel electrophoresis, Salmonella spp.

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Identification and classification of honey's authenticity by attenuated total reflectance Fourier-transform infrared spectroscopy and chemometric method

Research (Published online: 23-08-2019)

19. Identification and classification of honey's authenticity by attenuated total reflectance Fourier-transform infrared spectroscopy and chemometric method

Muhamad Sahlan, Seffiani Karwita, Misri Gozan, Heri Hermansyah, Masafumi Yohda, Young Je Yoo and Diah Kartika Pratami

Veterinary World, 12(8): 1304-1310

ABSTRACT

Background and Aim: The authentication of honey is important to protect industry and consumers from such adulterated honey. However, until now, there has been no guarantee of honey's authenticity, especially in Indonesia. The classification of honey is based on the bee species (spp.) that produces it. The study used honey from sting bee Apis spp. and stingless bee Tetragonula spp. based on the fact that the content off honey produced between them has differences. Authenticating honey with currently available rapid detection methods, such as 13C nuclear magnetic resonance analysis, is costly. This study aimed to develop an inexpensive, fast, precise, and accurate classification method for authenticating honey.

Materials and Methods: In this study, we use attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy with wavelengths ranging between 550 and 4000 cm-1 as an alternative analysis method, which is relatively less expensive. The spectra of authentic and fake honey samples were obtained using ATR-FTIR and plotted using chemometric discriminant analysis. The authentic honey samples were acquired from a local Indonesian breeder of honey bees, while the fake honey samples were made from a mixture of water, sugar, sodium bicarbonate, and authentic honey. Data were collected using Thermo Scientific's OMNIC FTIR software and processed using Thermo Scientific's TQ Analyst software.

Results: Our method effectively classified the honey as authentic or fraudulent based on the FTIR spectra. To authenticate the honey, we formed two classes: Real honey and fake honey. The wavelengths that can best differentiate between these two classes correspond to four regions: 1600-1700 cm-1; 1175-1540 cm-1; 940-1175 cm-1; and 700-940 cm-1. Similarly, for classification purpose, we formed two classes: Apis spp. and Tetragonula spp. The wavelength region that can best classify the samples as belonging to the Apis spp. or Tetragonula spp. class is explicitly within the range of 1600-1700 cm-1.

Conclusion: This study successfully demonstrated a method to rapidly and accurately classify and authenticate honey. ATR-FTIR is a useful tool to test the authenticity of honey.

Keywords: Apis spp., attenuated total reflectance Fourier transform infrared, discriminant, spectrum, Tetragonula spp.

Effect of the pH in the enrichment of X or Y sex chromosome-bearing sperm in bovine

Research (Published online: 23-08-2019)

18. Effect of the pH in the enrichment of X or Y sex chromosome-bearing sperm in bovine

Nidhi P. Raval, Tejas M. Shah, Linz-Buoy George and Chaitanya G. Joshi

Veterinary World, 12(8): 1299-1303

ABSTRACT

Background and Aim: Studies have shown that the pH of the vagina during the course of fertilization may influence the migration of X- and Y-bearing spermatozoa and thus leading to skewness in the sex of the offspring. Hence, this study was carried out to check the effect of the pH in the enrichment of X or Y sex chromosome-bearing sperm in bovine (Bos indicus).

Materials and Methods: To check the effect of pH in the enrichment of X or Y sex chromosome-bearing sperm in bovine, we used buffers of various pH ranging from 5.5 to 9.0 for swim-up procedure of sperm sample and collected upper and bottom fraction from the same buffer and checked the abundance of X- and Y-bearing spermatozoa by droplet digital polymerase chain reaction using X- and Y-chromosome-specific DNA probe.

Results: The abundance of X- and Y-bearing spermatozoa was not differed significantly in either of the fraction collected.

Conclusion: Thus, it appears to be unlikely that an immediate impact of pH on sperm can be a solitary impact on the sex of offspring in bovine.

Keywords: droplet digital polymerase chain reaction, spermatozoa, swim-up.

Field safety and efficacy of a unique live virus vaccine for controlling avian encephalomyelitis and fowlpox in poultry

Research (Published online: 23-08-2019)

17. Field safety and efficacy of a unique live virus vaccine for controlling avian encephalomyelitis and fowlpox in poultry

Girish Sarma, Barry A. Kersting and Gary Spina

Veterinary World, 12(8): 1291-1298

ABSTRACT

Background and Aim: Infection of commercial poultry with avian encephalomyelitis (AE) and fowlpox (FP) virus causes heavy economic loss in endemic areas. Although vaccines are routinely used to control these two diseases, the problem still persists almost all over the world. This study aimed to evaluate safety and efficacy of a unique AE + FP + pigeon pox (PP) live virus vaccine in layer-type chickens under both laboratory and field conditions.

Materials and Methods: The study was conducted using 289 specific-pathogen-free (SPF) chickens under the laboratory conditions and 185,648 commercial layer-type chickens under field conditions. In two consecutive laboratory trials, 8-week-old SPF chickens were vaccinated with the AE + FP + PP live virus vaccine through wing web route and challenged against virulent strains of FP and AE viruses at 3-week post-vaccination (WPV). Challenged chickens were observed for disease protection for 10-21 days. For field safety trials, commercial layer-type chickens in three different geographical areas in the USA were vaccinated with the AE + FP + PP vaccine and observed daily up to 21 days for vaccine "take". adverse reactions, and mortality.

Results: The vaccine was found safe and efficacious under both laboratory and field conditions. Vaccine "take" and protection against FP challenge were 100%. Average protection against AE challenge was 97%. Mean AE enzyme-linked immunosorbent assay (ELISA) antibody titer in the field vaccinated chickens was >1200 at 10 WPV. Average daily post-vaccination mortality in the field vaccinated chickens was 0.04%. So far, more than 400 million chickens in the USA have been vaccinated with this vaccine. No vaccine-associated adverse reactions, other safety issues, or immunity breakdown cases in the vaccinated flocks due to field virus infection have been reported.

Conclusion: This unique vaccine containing AE, FP, and PP viruses in a single preparation was found to be safe and efficacious in controlling the diseases caused by the virulent field strains of AE and FP. Besides being safe and efficacious, this vaccine also offered distinct advantages over the traditional vaccination practices in controlling these two diseases in poultry.

Keywords: avian encephalomyelitis, efficacy, field safety, fowlpox, live virus vaccine, pigeon pox, protection.

Knowledge and behavior of cattle and sheep owners and herders regarding foot-and-mouth disease in Northern Algeria

Research (Published online: 21-08-2019)

16. Knowledge and behavior of cattle and sheep owners and herders regarding foot-and-mouth disease in Northern Algeria

Ratiba Baazizi, Nora Mimoune, M'Hamed El Mokhefi, Muslim Raza, Amina Chahed and Tanveer Hussain

Veterinary World, 12(8): 1285-1290

ABSTRACT

Background and Aim: Foot-and-mouth disease (FMD) has been occurring in Algeria since 2014, when an outbreak was announced in Setif, a district in the eastern region of the country. The problem was apparently resolved with the help of vaccination. However, in 2015, 2016, and 2018, FMD recurred. The veterinary authorities and media educated breeders on how to recognize the clinical signs and how to report the disease. This study aimed to evaluate the knowledge and recognition of FMD by farmers and breeders. Moreover, an assessment of the behavior of cattle and sheep owners and herders following FMD cases is examined.

Materials and Methods: A cross-sectional survey was conducted from June to October 2018 to evaluate the perception of cattle and sheep owners and breeders regarding FMD in the Northern regions of Algeria, using questionnaires.

Results: One hundred questionnaires were distributed; 71 were collected. Data showed that all the responders claimed to know about the disease, while more than half of the owners/herders claimed that they knew the clinical symptoms of FMD and mentioned fever, hypersalivation, lameness, and vesicles. Fewer than half (42%) (30/71) took some measures to prevent the disease, while more than half (58%) (41/71) did not take any measures in 2018. No one claimed to have reported the disease to authorities in 2018, while more than half had done so in 2014.

Conclusion: It appears that experienced farmers recognized the clinical signs of FMD, while an academic background was not conclusively necessary for the identification of the clinical signs of the disease. Concerning the assessment of risk-associated behavior in the event of FMD occurrence, the responses of the breeders were not significantly different from those of risk-associated behaviors in the event of an epidemic. Farmers and breeders expressed similarity in terms of communicating the appearance of the disease in their livestock; the majority of them seemed to be aware of the importance of reporting the disease to local authorities, especially in 2014, when the disease first occurred. This behavior is encouraged by refund and technical assistance policies by the veterinary authorities, but in 2018, no disease was reported due to fear of slaughtering and economic loss.

Keywords: cattle and sheep farmers, clinical signs, foot-and-mouth disease, knowledge and seniority, survey.

Risk factors associated with viral nervous necrosis in hybrid groupers in Malaysia and the high similarity of its causative agent nervous necrosis virus to reassortant red-spotted grouper nervous necrosis virus/striped jack nervous necrosis virus strains

Research (Published online: 21-08-2019)

15. Risk factors associated with viral nervous necrosis in hybrid groupers in Malaysia and the high similarity of its causative agent nervous necrosis virus to reassortant red-spotted grouper nervous necrosis virus/striped jack nervous necrosis virus strains

Nurshuhada Ariff, Azila Abdullah, Mohamed Noor Amal Azmai, Najiah Musa and Sandra Catherine Zainathan

Veterinary World, 12(8): 1273-1284

ABSTRACT

Background and Aim: Viral nervous necrosis (VNN) is a serious disease of several marine fish species. VNN causes 100% mortality in the larval stages, while lower losses have been reported in juvenile and adult fish. This study aimed to detect the occurrence of VNN while identifying its associated risk factors and the genotypes of its causative agent in a hybrid grouper hatchery in Malaysia.

Materials and Methods: A batch of newly hatched hybrid grouper fry (Epinephelus fuscoguttatus X Epinephelus lanceolatus) were followed from the larval stage to market size. Samples of the hybrid groupers, water, live feed, and artificial fish pellets were collected periodically from day 0 to 180 in the hybrid grouper hatchery. Reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR amplifications were carried out on VNN-related sequences. The phylogenetic tree including the sampled causative agent of VNN was inferred from the coat protein genes from all known Betanodavirus species using Molecular Evolutionary Genetics Analysis (MEGA). Pearson's correlation coefficient values were calculated to determine the strength of the correlation between the presence of VNN in hybrid grouper samples and its associated risk factors.

Results: A total of 113 out of 146 pooled and individual samples, including hybrid grouper, water, and artificial fish pellet samples, demonstrated positive results in tests for the presence of VNN-associated viruses. The clinical signs of infection observed in the samples included darkened skin, deformation of the backbone, abdominal distension, skin lesions, and fin erosion. VNN was present throughout the life stages of the hybrid groupers, with the first detection occurring at day 10. VNN-associated risk factors included water temperature, dissolved oxygen content, salinity, ammonia level, fish size (adults more at risk than younger stages), and life stage (age). Detection of VNN-associated viruses in water samples demonstrated evidence of horizontal transmission of the disease. All the nucleotide sequences found in this study had high nucleotide identities of 88% to 100% to each other, striped jack nervous necrosis virus (SJNNV), and the reassortant strain red-spotted grouper NNV/SJNNV (RGNNV/SJNNV) isolate 430.2004 (GenBank accession number JN189932.1) (n=26). The phylogenetic analysis showed that quasispecies was present in each VNN-causing virus-positive sample, which differed based on the type of sample and life stage.

Conclusion: This study was the first to confirm the existence of a reassortant strain (RGNNV/SJNNV) in hybrid groupers from Malaysia and Southeast Asia. However, the association between the mode of transmission and the risk factors of this virus needs to be investigated further to understand the evolution and potential new host species of the reassortant strain.

Keywords: epidemiology, quasispecies, reassortant, red-spotted grouper nervous necrosis virus-striped jack nervous necrosis virus, viral nervous necrosis.

Evaluation of hepatobiliary ultrasound scores in healthy dogs and dogs with liver diseases

Research (Published online: 17-08-2019)

14. Evaluation of hepatobiliary ultrasound scores in healthy dogs and dogs with liver diseases

Sathidpak Nantasanti Assawarachan, Piyathip Chuchalermporn, Phudit Maneesaay and Naris Thengchaisri

Veterinary World, 12(8): 1266-1272

ABSTRACT

Background and Aim: Ultrasonography is the first-line method for examining the canine liver. Hepatic ultrasound scoring systems are widely described in human medicine, yet there is no information on the use of semi-quantitative ultrasound scoring systems in canine liver diseases. This study aimed to evaluate the hepatobiliary ultrasound scores between physically healthy dogs and dogs with primary liver diseases confirmed by clinical, biochemical, and histological parameters. We also evaluated the putative correlations between ultrasound scores and ALT or ALP levels. Moreover, the severity of ultrasound scoring and fold changes in liver enzymes was also evaluated.

Materials and Methods: A cross-sectional study design was conducted to compare the results of the six different parameters (liver surface, echogenicity of parenchyma, nodularity of parenchyma, gallbladder wall thickness, amount of gall sludge, and visibility of bile duct) of ultrasound scores between dogs with and without liver disease.

Results: Our results showed that 17.4%, 88.2%, and 100% of dogs with liver diseases were identified according to the ultrasound severity classified as mild (total score 0-2), moderate (total score 3-5), and severe (total score 6-12). Approximately 30% of patients with chronic hepatitis, the most common canine liver disease, presented with normal or mild ultrasound score category, whereas most of the patients with vacuolar hepatopathy and steroid-induced hepatopathy due to secondary reactive changes had moderate-to-severe ultrasound score category. There were 75% of patients with tumor and 80% of patients with hepatic fibrosis that were identified with severe ultrasound score category. Dogs with moderate-to-severe ultrasound scores had significant liver enzyme elevation (both alanine aminotransferase [ALT] and alkaline phosphatase [ALP]) compared to those of dogs with mild ultrasound scores. Ultrasound score was moderately associated with ALT and highly associated with ALP levels (p=0.553 and p=0.730, respectively).

Conclusion: Our semi-quantitative, simplified ultrasonographic scoring system may have potential to be used as a screening tool to detect some groups of liver diseases.

Keywords: diagnosis, dogs, hepatic disease, ultrasound score.

Effects of a standardized 40% ellagic acid pomegranate (Punica granatum L.) extract on seminiferous tubule histopathology, diameter, and epithelium thickness in albino Wistar rats after heat exposure

Research (Published online: 17-08-2019)

13. Effects of a standardized 40% ellagic acid pomegranate (Punica granatum L.) extract on seminiferous tubule histopathology, diameter, and epithelium thickness in albino Wistar rats after heat exposure

Budi Utomo, Nurfitri Rahmah Daningtia, Gandul Atik Yuliani and Wiwik Misaco Yuniarti

Veterinary World, 12(8): 1261-1265

ABSTRACT

Background and Aim: It has long been known that the spermatogenic tissue is very sensitive to temperatures higher than its physiologic temperature and causing cessation of activity and resulting in sterility. This study investigated the effect of a standardized 40% ellagic acid extract of pomegranate on the histopathology, diameter, and epithelial thickness of seminiferous tubules in albino rats exposed to heat.

Materials and Methods: Twenty-five male albino Wistar rats were randomized at 7-8 months of age to five treatment groups. Group C was not treated; Group T0 was treated with 0.5% of Na carboxymethyl cellulose (CMC) 2 ml/day and exposed to heat. T1, T2, and T3 were treated with 75, 150, and 300 mg/kg/day of a standardized 40% ellagic acid extract of pomegranate (Punica granatum L.), respectively. The animals were orally administered Na CMC or pomegranate extract and were exposed to sunlight for 15 min at 40°C-42°C for 14 days. The animals were sacrificed on day 15 and the testes were removed for histological evaluation and measurement of seminiferous tubule diameter and epithelium thickness.

Results: Our results showed that 17.4%, 88.2%, and 100% of dogs with liver diseases were identified according to the ultrasound severity classified as mild (total score 0-2), moderate (total score 3-5), and severe (total score 6-12). Approximately 30% of patients with chronic hepatitis, the most common canine liver disease, presented with normal or mild ultrasound score category, whereas most of the patients with vacuolar hepatopathy and steroid-induced hepatopathy due to secondary reactive changes had moderate-to-severe ultrasound score category. There were 75% of patients with tumor and 80% of patients with hepatic fibrosis that were identified with severe ultrasound score category. Dogs with moderate-to-severe ultrasound scores had significant liver enzyme elevation (both alanine aminotransferase [ALT] and alkaline phosphatase [ALP]) compared to those of dogs with mild ultrasound scores. Ultrasound score was moderately associated with ALT and highly associated with ALP levels (p=0.553 and p=0.730, respectively).

Conclusion: Compared with controls, administration of 300 mg/kg/day of a standardized 40% ellagic acid extract of P. granatum L. for 14 days increased seminiferous tubule diameter and epithelium thickness in albino Wistar rats exposed to heat.

Keywords: diameter, epithelium thickness, heat, pomegranate extract, seminiferous tubule.

Intestinal and hepatic coccidiosis among rabbits in Yogyakarta, Indonesia

Research (Published online: 15-08-2019)

12. Intestinal and hepatic coccidiosis among rabbits in Yogyakarta, Indonesia

Penny Humaidah Hamid, Sigit Prastowo and Yuli Purwandari Kristianingrum

Veterinary World, 12(8): 1256-1260

ABSTRACT

Background and Aim: The attention to rabbit meat production in Indonesia is comparatively less to other farm animals such as cattle and poultry industries. However, future prospect of rabbit to be seriously industrialized seemed quite promising due to rabbit is highly productive and has short reproduction cycle as well as generation interval. One of the diseases infecting many rabbits is coccidiosis caused by protozoan parasite, Eimeria spp. The infectious stage of Eimeria spp. presents ubiquitously in the environment and increases the risk of parasite transmission. Preventive methods such as vaccination are not yet fully developed, while sporadic treatment is not efficiently reduce the cases. In this study, Eimeria spp. infecting rabbits in Yogyakarta Province, Indonesia, were investigated with the aim for precise diagnosis to determine targeted treatment and as a baseline epidemiological data from rabbit in Indonesia.

Materials and Methods: Sample collection was performed randomly for 3 months, from March 2017 to May 2017 and covered areas in Yogyakarta, Indonesia. A total of 750 samples were collected. Eimeria species identification was determined morphologically from the samples after sporulation in 2.5% potassium dichromate by COCCIMORPH.

Results: Ten species of Eimeria spp. were identified in this study from the positive samples (527/750; 70.3%). Eimeria flavescens was present in 80% of the positive samples, Eimeria coeciola in 78%, Eimeria perforans in 61%, Eimeria exigua in 37%, Eimeria media in 33%, Eimeria stiedae in 31%, Eimeria irresidua in 12%, Eimeria magna in 11%, Eimeria intestinalis in 10%, and Eimeria piriformis in 10%. Coinfection as noted in 80% of the positive samples with 2-6 species in a specimen. E. flavescens and E. coeciola were the most prevalent among all Eimeria spp. (p=0.0001).

Conclusion: Eimeria spp. is detected in high prevalence among rabbit in Yogyakarta, Indonesia, with commonly occurs in mixed infections. In this paper, we describe Eimeria spp. that are circulating in Indonesia and present it as updated information to farmers and veterinarians. To the best of our knowledge, we provided the first information about rabbit coccidiosis in Indonesia.

Keywords: coccidiosis, Indonesia, rabbit.

Studies on immune response to Newcastle disease virus in broiler chickens fed with Lactobacillus reuteri PIA16 isolated from the gut of indigenous chicken of Assam, India

Research (Published online: 15-08-2019)

11. Studies on immune response to Newcastle disease virus in broiler chickens fed with Lactobacillus reuteri PIA16 isolated from the gut of indigenous chicken of Assam, India

Gaichamdinliu Gonmei, Deben Sapcota, Girin Kumar Saikia, Pankaj Deka, Joga Dev Mahanta, Niranjan Kalita, Bibeka Nanda Saikia and Jitendra Kumar Talukdar

Veterinary World, 12(8): 1251-1255

ABSTRACT

Background and Aim: The chicken gut harbors microflora which impacts the health, production performance and immune response against pathogens. Assam local chickens reared under natural conditions are known to possess high immunocompetence which may be attributable to its gut microbiota make-up. This study aimed to investigate the individual effect of two strains of Lactobacillus reuteri PIA16 isolated separately from cecum and jejunum of Assam indigenous chicken on the immunity of broiler chickens against Newcastle disease virus (NDV) when fed singly and in combination with a prebiotic.

Materials and Methods: A total of 240 birds (48 per group) were vaccinated with Lasota strain of NDV on the 5th and 21st day of age. Blood samples were collected before and after immunization against ND for the detection of humoral antibody response by hemagglutination inhibition test. The cell-mediated immune (CMI) response was estimated through response to phytohemagglutinin-P (PHA-P) and expressed as web index.

Results: A significant influence on the immune response to NDV was observed in all the L. reuteri PIA16 as well as mannan oligosaccharide (MOS) supplemented groups revealing higher antibody titer than the control counterpart. The CMI response revealed a better cutaneous basophilic hypersensitivity response to PHA-P in the treated groups than the control.

Conclusion: Enhancement in immunity was perceived in the broilers fed with L. reuteri PIA16 and in combination with MOS due to the stimulation of the host's humoral and CMI response by the probiotics and prebiotics used.

Keywords: hemagglutination inhibition, Lactobacillus reuteri PIA16, mannan oligosaccharide, Newcastle disease, phytohemagglutinin-P, probiotics.

Epidemiological study of sausage in Algeria: Prevalence, quality assessment, and antibiotic resistance of Staphylococcus aureus isolates and the risk factors associated with consumer habits affecting foodborne poisoning

Research (Published online: 15-08-2019)

10. Epidemiological study of sausage in Algeria: Prevalence, quality assessment, and antibiotic resistance of Staphylococcus aureus isolates and the risk factors associated with consumer habits affecting foodborne poisoning

Amina Hachemi, Safia Zenia, Mohamed Fatih Denia, Meryem Guessoum, Mohamed Mehdi Hachemi and Khatima Ait-Oudhia

Veterinary World, 12(8): 1240-1250

ABSTRACT

Aim: The first aim was to assess the quality and determine the prevalence and antibiotic susceptibility of Staphylococcus aureus contamination of raw sausage sold in ten municipalities in the Northeast of Algeria. Second, a consumer sausage purchasing survey was designed to investigate potential risk factors that have a significant association with the occurrence of foodborne poisoning among sausage consumers' behavior and its relationship with independent variables.

Materials and Methods: A total of 230 butcheries from ten departments (Daira) of Algiers with more than 40 municipalities were included randomly in these studies to collect raw sausage samples and to distribute 700 structured questionnaires to meat consumers. Our two studies were conducted at the same time, between June 2016 and April 2018. Sausage samples were taken once per butchery to estimate the prevalence of S. aureus contamination and therefore deduct the quality assessment of raw sausage (Merguez) sold in Algiers, Algeria. All isolated strains were tested for their antimicrobial resistance. Furthermore, questionnaires were distributed and used to collect information on various aspects of sausage consumption and foodborne disease. The data collected were analyzed with different statistical approaches, such as the Chi-square test and the odds ratio (OR) univariable logistic model. All the risk factors were analyzed by studying their association with the occurrence of consumers who claimed to have food poisoning after consuming sausage.

Results: The overall prevalence of S. aureus contamination from sausages was 25.22% (n=58/230). Over 83.33% of strains showed resistance to at least one of the antibiotics tested. The most important was for tetracycline (58%) followed by fosfomycin (33%), penicillin G (25%), and oxacillin (36%). Moreover, the multiple antibiotic resistance (MAR) index include 20 profiles with MAR >0.2. Out of the 440 meat consumers, 22.16% revealed having food poisoning after sausage consumption. The risk factors recorded were: Consumption outside of home (24.30%, OR=1.769, p=0.040), during the summer season (24.30%, OR=1.159) and during lunch (26.50%, OR=1.421).

Conclusion: Our study highlights a high prevalence of S. aureus contamination in Merguez, especially in some departments of Algiers, and the high multidrug resistance of S. aureus isolates against tetracycline and oxacillin; thus, S. aureus contamination in sausage is considered a potential risk to public health. Therefore, to reduce and prevent the spread of resistant strains, robust management and monitoring of antibiotic use should be established. Therefore, it is necessary to improve the sanitation conditions and education regarding personal hygiene and change certain consumption habits of Algerian consumers to ensure food safety. Finally, it can be concluded that the application of the HACCP system is essential either in butcheries producing sausage and/or slaughterhouses. From this perspective, studies might be performed to characterize Staphylococcus spp. and S. aureus to investigate their virulence factors.

Keywords: consumers, quality assessment, risk factors, sausages, Staphylococcus aureus.