Detection of canine parvovirus type 2c (CPV-2c) in Palestine.

INTRODUCTION: The objective of the present study was to report, for the first time, the presence of canine parvovirus type 2c (CPV-2c) in domesticated dogs with acute gastroenteritis and to characterize the antigenic variants circulating in Palestine. METHODOLOGY: A veterinary clinical-based epidemiological study was carried out between December 2022 and April 2023. Fifty fecal samples were collected from dogs with gastroenteritis and screened for CPV-2 infection by polymerase chain reaction. The distribution of positive cases according to various epidemiological factors was studied. Partial sequencing of the viral protein 2 (VP2) gene was performed for the analysis of CPV-2 variants. RESULTS: Most of the investigated samples (60%; n = 50) during the study period were found positive for CPV-2 infection. There was no difference in the distribution of positive cases of CPV-2 infection based on age group, gender, location, and vaccination status. The analysis of nucleotide and amino acid sequences from amplified products, as well as phylogenetic analysis, revealed the presence of CPV-2c clustered with Asian CPV-2c variants. CONCLUSIONS: In summary, this study represents the initial genetic analysis of CPV-2 present in Palestinian dogs with gastroenteritis and provides evidence that confirms the existence of the CPV-2c variants. To determine the prevailing CPV-2 variant associated with the infection, it is crucial to conduct further sequence analysis using large populations of both domestic and wild canines.

First report of mobile colistin resistance gene mcr-1 in avian pathogenic Escherichia coli isolated from turkeys in the Gaza Strip, Palestine.

Background and Aim: Colistin is used to treat avian pathogenic Escherichia coli (APEC), a microorganism that affects turkey meat production in the Gaza Strip and worldwide. However, the recent emergence of plasmid-borne mobile colistin resistance (mcr) genes in pathogenic E. coli strains is a serious antimicrobial resistance (AMR) challenge for both human and animal health. In December 2018, colistin was banned as a veterinary antimicrobial in the Gaza Strip. This study aimed to detect and track the prevalence of colistin-resistant APEC isolated from turkey flocks in the Gaza Strip. Materials and Methods: This study investigated 239 APEC isolates from turkey flocks in the Gaza Strip between October 2018 and December 2021 (at 6-month intervals). The colistin-resistant APEC strains were detected using the broth microdilution method. The mcr-1 gene was identified using a polymerase chain reaction. Results: The overall colistin resistance among the isolated APECs was 32.2% during the study period. The average resistance in the first interval was 37.5%, which significantly decreased to 9.3% in the last interval. Among the 77 phenotypically resistant isolates, 32.4% were positive for mcr-1. The average abundance of mcr-1 in the first interval was 66.6%, which decreased to 25% in the last interval. Conclusion: To the best of our knowledge, this is the first study reporting the presence of the mcr-1 gene among the APEC isolates from turkeys in the Gaza Strip. Banned veterinary use of colistin significantly decreased the percentage of resistant APEC isolates from turkeys in Gaza Strip. Further studies are needed to investigate other colistin resistance genes and track the emergence of AMR.

Occurrence and phylogenetic analysis of fowl adenovirus E in broiler flocks from Gaza Strip Palestine.

INTRODUCTION: Inclusion-body hepatitis (IBH) and hydropericardium syndrome (HPS) are highly infectious diseases caused by fowl adenoviruses (FAdVs). IBH and HPS cause major economic losses in poultry production. IBH is caused by multiple FAdV serotypes such as FAdV-11, FAdV8a, and FAdV8b; while HPS is mainly caused by the FAdV-4 serotype. In 2018, we detected FAdVs in West Bank - Palestine for the first time. This study aims to monitor the emergence of new FAdVs in broiler farms in Gaza Strip-Palestine in 2022. METHODOLOGY: The clinical signs, necropsy, and histopathological findings associated with IBH in the suspected birds were recorded. Polymerase chain reaction (PCR) was performed using primers matching the virus-encoded L1 loop of the hexon gene. The sequences of the L1 loop were analyzed and a phylogenetic tree was constructed and compared with the related FAdV field isolates and reference strains from different regions of the world available in GenBank. RESULTS: The infected broiler displayed FAdVs-induced clinical symptoms and pathological lesions with mortality rates ranging from 20-46%. The L1 loop sequences from the infected flocks were submitted to GenBank with accession numbers ON638995, ON872150, and ON872151. The identified L1 loop gene has high nucleotide homology (96.7-97.9%) to the highly pathogenic FAdV E serotype 8b strain FAdV_isolate_04-53357-122_Canada_2007 (GenBank: EF685489) and 94.5-94.6% to FAdV_10_Belgium_2010 isolate 11-15941 (GenBank: AF339924.1). Furthermore, the phylogenetic analysis indicated that they belong to FAdV-E serotype 8b. CONCLUSIONS: Our study reports the emergence of FAdV-E causing IBH disease in broiler chickens for the first time in Gaza in Palestine.

Pathological and molecular study of kidneys in apparently healthy cattle and sheep with special reference to Leptospira species in central and northern Jordan.

Background and Aim: Renal pathological conditions can cause significant economic losses for livestock owners. This study investigated and described the gross pathology and histopathology of lesions found in the kidneys of sheep and cattle, with particular attention to the presence of Leptospira spp. Materials and Methods: Three hundred and sixty kidneys from apparently healthy sheep and cattle (208 and 152, respectively) were collected from slaughterhouses in Jordan for gross and histopathological examinations, multiplex polymerase chain reaction (PCR) analysis, and gene sequencing of Leptospira spp. Results: Histopathological analysis of the samples revealed the following conditions: interstitial nephritis (4.44%), glomerulonephritis (4.16%), acute tubular necrosis (17.22%), and renal congestion (4.72%). In addition, in 26.9% of the samples, hyaline material was observed in the interstitium of the cortex and medulla. The PCR results revealed that 83 (23.1%) of the 360 samples were positive for Leptospira spp. 16S rRNA, 42/152 (28%) of cattle, and 41/208 (20%) of sheep. Four samples (two sheep and two cattle) were sequenced and deposited in GenBank with accession numbers from OL701310 to OL701313. Basic Local Alignment Search Tool search and nucleotide similarities between OL701311 and OL701313 resulted in the highest similarities with different Leptospira borgpetersenii strains, whereas OL701310 showed the highest nucleotide similarity (99.2%) with the Leptospira interrogans strain. Similarly, phylogenetic analysis revealed that OL701311 to OL701313 clustered together with different serovars of L. borgpetersenii, whereas OL701310 clustered with the L. interrogans clade. Conclusion: This is the first study to reveal a close association between pathogenic Leptospira spp. and kidney disorders in Jordanian cattle and sheep. These findings may help expand the current understanding of the causes and mechanisms of renal disease in cattle and sheep and contribute to developing more effective prevention and treatment programs.

The Impact of Thyme and Oregano Essential Oils Dietary Supplementation on Broiler Health, Growth Performance, and Prevalence of Growth-Related Breast Muscle Abnormalities.

The objective of this study was to investigate the effects of thyme and oregano essential oils (as growth promotors), individually and in combination, on the health, growth performance, and prevalence of muscle abnormalities in broiler chickens. Six hundred day-old Cobb 500 hybrid chickens were randomized into four dietary treatment groups with three replicates each. Chicks in the control group (C) received a basal diet, while the experimental treatment groups received basal diets containing 350 mg/kg of thyme oil (T1), 350 mg/kg of oregano oil (T2), and 350 mg/kg of thyme and oregano oil (T3). Growth performance parameters were evaluated at 14, 28, and 42 days. The broilers in treatments T1 and T2 had significantly higher body weights than the control group. The feed conversion ratio was the lowest in chicks who received oregano oil, followed by those fed thyme oil. The overall prevalence of growth-related breast muscle abnormalities (including white striping and white striping combined with wooden breast) in groups receiving essential oils (T1, T2, and T3) was significantly higher than in the control group (C). The thyme and oregano oil diets showed no significant differences in antibody titers against Newcastle disease or interferon-γ (INF-γ) serum levels. In conclusion, thyme and oregano oils had a positive impact on the growth performance of broiler chickens but increased the incidence of growth-related breast muscle abnormalities.

No evidence of Rift Valley fever antibodies in veterinarians and sheep in Northern Palestine.

Background and Aim: Rift Valley fever virus (RVFV) is a vector-borne virus that causes RVF in humans and ruminants. The clinical symptoms in humans and animals are non-specific and often misdiagnosed, but abortions in ruminants and high mortality in young animals are characteristic. Since the initial outbreak in the Rift Valley area in Kenya, the disease has spread to most African countries and the Middle East. The presence and epidemiological status of RVFV in humans and animals in Palestine are unknown. This study aimed to investigate the presence and risk factors for RVF seroprevalence in veterinarians, as occupational hazard professionals, and sheep, as highly susceptible animals, in Northern Palestine. Materials and Methods: A cross-sectional study was conducted. Data and blood samples of 280 Assaf sheep and 100 veterinarians in close occupational contact with sheep were collected between August and September 2020 using an indirect enzyme-linked immunosorbent assay. Results: No evidence of RVF antibodies was found in any human or animal sample. Conclusion: Our results suggest that RVFV has not circulated in livestock in Northern Palestine, yet. Surveillance and response capabilities and cooperation with the nearby endemic regions are recommended. The distribution of competent vectors in Palestine, associated with global climate change and the role of wild animals, might be a possible route for RVF spreading to Palestine from neighboring countries.

In vitro activity of selected natural products against Eimeria tenella sporozoites using reproduction inhibition assay.

Eimeria tenella is the causative agent of cecal coccidiosis in poultry characterized by weight loss, hemorrhagic diarrhea, and high mortality rates. Research into herbal candidates with possible anticoccidial activity has increased lately. As an alternative to animal experiments, an in vitro reproduction inhibition assay (RIA) was previously designed to determine the sensitivity of E. tenella isolates against ionophores. In this study, the RIA was used to test the anticoccidial activity of nutmeg oil, cinnamon oil, and glabridin. The concentration of nutmeg oil used in this study ranged between 1.1 and 139.1 µg/ml. Nutmeg oil exhibited a moderate in vitro inhibitory activity ranging from 35.5 to 49.5%. In contrast, no inhibitory effect was detected when incubating E. tenella sporozoites for 24 h with cinnamon oil at concentrations of 0.3 to 80.5 µg/ml. Glabridin (0.08-41.7 µg/ml) prevented the replication of sporozoites at a rate of 14.1 to 81.7% of inhibition. The calculated minimum concentrations of glabridin needed to inhibit parasite replication by 75%, 50%, and 30% (MIC75, MIC50, and MIC30) were 21.43 µg/ml, 5.28 µg/ml, and 0.96 µg/ml, respectively. Further studies to assess the in vitro efficacy of glabridin were performed by studying mRNA gene expression of stress-induced protein genes (HSP-70, NADPH, and EtPP5) after exposure of E. tenella sporozoites to glabridin at MIC75 for 0.5 h, 1 h, 2 h, and 4 h (a time-dependent experiment). Moreover, a dose-dependent experiment was performed using glabridin at a concentration matching MIC75, MIC50, and MIC30 for 24 h. In the time-dependent experiment, a significant (p < 0.05) increase of expression in NADPH and EtPP5 were detected after 4 h of incubation with glabridin at a concentration of 21.43 µg/ml. The dose-dependent experiment exhibited a gradual increase of expression in all studied genes, which indicates stress imposed on E. tenella sporozoites by glabridin. In our hands, RIA was suitable to assess the anticoccidial activity exhibited by the tested natural products as a precursor to in vivo studies which will help in the identification of novel anticoccidial candidates.

Seroprevalence and risk factors of West Nile virus infection in veterinarians and horses in Northern Palestine.

BACKGROUND AND AIM: West Nile fever (WNF) is a neurotropic, mosquito-borne disease affecting humans and domesticated animals, caused by a member of the genus Flavivirus. Over the last decades, this virus has been responsible for several cases of illness in humans and animals. The current epidemiological status of WNF in horses is insufficient, and in veterinarians, as an occupational hazard is unknown. This study aimed to investigate and determine the seroprevalence and risk factors for WNF in veterinarians and horses in Palestine. MATERIALS AND METHODS: In this study, serum samples from 100 veterinarians and 87 horses were collected between August 2020 and September 2020 from different cities of Northern Palestine. West Nile virus (WNV) antibodies were detected using an enzyme-linked immunosorbent assay. RESULTS: Our results showed that 60.9% of the horse serum samples were positive in all investigated cities. In horses, location is a risk factor for the seropositivity for WNF, whereas age, sex, breed, and intended use of the horses, were not associated with increased WNF seropositivity. In veterinarians, 23.0% of the serum samples were positive. Positive samples were detected in all locations, age groups, experience length, and work sectors. However, the seropositivity for WNF was not influenced by these variables. CONCLUSION: The results revealed that WNV circulates in most regions of Palestine. Our results will help determine the risk of infection in animals and humans and control WNV transmission. Surveillance studies on humans, vectors, and animals are needed to better define endemic areas.

Phylogenetic analysis and pathological characterization of fowl adenovirus isolated during inclusion body hepatitis outbreak in Tubas, Palestine.

Fowl adenoviruses (FAdVs) associated with inclusion body hepatitis (IBH) was identified in commercial broiler chickens in Palestine. Investigated birds showed primary clinical signs and lesions of IBH including growth retardation, congested and enlarged liver with necrosis, petechial hemorrhage and basophilic intra-nuclear inclusion bodies. The mortality rate was from 15.00%. The FAdV was detected and sequenced in the liver samples of infected chicken by polymerase chain reaction using hexon gene-specific primers. Phylogenetic analysis revealed that FAdVs belong to FAdV-D serotype 10, clustered within the European highly pathogenic isolates. The highest nucleotide sequence similarity was 99.48% with highly pathogenic FAdV-D serotype 10 detected from infected chicken in Poland (GenBank: LN907532.1) and FAdV-D from infected chicken in Sweden (GenBank: HE961828.1). The lowest similarity was 93.46% with Canadian FAdV-D (GenBank: EF685576.1). In conclusion, this is the first report describing the presence of IBH revealing that the causative virus is closely similar to the highly pathogenic FAdV-D serotype 10 of IBH in broiler chickens in Palestine.

Occupational Exposure Assessment and Seroprevalence of Brucella Specific Antibodies Among Veterinarians in the Northern Palestine.

Background: Brucellosis is one of the most popular zoonosis in the world caused by bacteria belonging to the genus Brucella. The disease is considered an occupational risk to persons dealing with animals and animal products. Brucellosis is endemic in livestock in Palestine. Yet, few studies investigated human brucellosis in Palestine. We aimed to estimate Brucella seropositivity among veterinary healthcare professionals in Northern Palestine, and to assess the associated risk factors. Methods: A cross-sectional study was conducted in four governorates in the Northern West Bank (Jenin, Nablus, Qalqylia, and Tulkarm). A sample of 100 veterinarians was collected. Participants were interviewed using a structured questionnaire to assess risk factors. Blood samples were collected to be screened for the presence of anti-Brucella IgG using the Enzyme-Linked Immunosorbent Assay (ELISA). Results: The seroprevalence of Brucellosis by ELISA was 76%. Risk factors included working in the public sector, dealing with animals' vaccination, longer period of exposure, and advancing in age. Conclusions: Brucellosis is a high-risk occupational disease among veterinarians. Its prevalence rate among veterinary healthcare workers in the Northern West Bank, Palestine was very high compared to neighboring countries and internationally.

A molecular and pathological study of peste des petits ruminants virus (PPRV) from field outbreaks in Palestine, 2017-2019.

Peste des petits ruminants (PPR) is a severe, highly contagious and fatal viral disease of small ruminants causing high economic losses. Therefore, the disease was targeted for eradication by 2030. The aim of this study was to investigate for the first time the molecular and pathological characterization of the circulating PPR virus (PPRV) in sheep and goat in Palestine. Samples were collected from suspected necropsy cases of sheep and goats during recent outbreaks in two provinces in Palestine between 2017 and 2019. In this study, severe PPR outbreaks occurred in sheep and goats causing typical lesions which include erosive and ulcerative stomatitis, bronchointerstitial pneumonia and severe enteritis. For the molecular investigation of PPRV, suspected animals were examined for the presence of PPRV by RT-PCR. PPRV genome was detected in all samples. Subsequently, two samples were used for N gene sequencing and phylogenetic analysis of PPRV isolates. The nucleotide sequence and phylogenetic analysis indicated that the Palestinian PPRV isolates were genetically clustered within the lineage IV isolates of the virus among populations of sheep and goats which most prevalent in Asia, the Middle East and recently Africa. Further analysis showed that the Palestinian isolates were closely related to those described in Turkey and Iraq, suggesting a common origin of PPRV isolates in the region. This information is critical to understand the molecular epidemiology of this disease in the region and helps to develop appropriate control measures for eradication of this disease.

Identification of the Receptor and Cellular Ortholog of the Marek's Disease Virus (MDV) CXC Chemokine.

Marek's disease virus (MDV) is a cell associated alphaherpesvirus that causes fatal lymphoma in chickens. One factor that plays a crucial role in MDV pathogenesis is the viral CXC chemokine vIL-8 that was originally named after chicken interleukin 8 (cIL-8). However, a recent study demonstrated that vIL-8 recruits B cells and a subset of T cells but not neutrophils, suggesting that vIL-8 is not a cIL-8 orthologue. In this study, we set to identify the cellular orthologues and receptor of vIL-8 using in silico analyses, binding and chemotaxis assays. Sequence and phylogenetic analyses of all chicken CXC chemokines present in the recently published chicken genome revealed that vIL-8 shares the highest amino acid similarity with the CXCL13L1 variant. To evaluate if vIL-8 and CXCL13L1 are also functional orthologues, we assessed their binding properties and chemotaxis activity. We demonstrated that both vIL-8 and CXCL13 variants bind B cells and subsets of T cells, confirming that they target the same cell types. In addition, the chemokines not only bound the target cells but also induced chemotaxis. Finally, we identified CXCR5 as the receptor of vIL-8 and CXCL13 variants and confirmed that the receptor is expressed on MDV target cells. Taken together, our data demonstrate the conservation of the receptor-ligand interaction between CXCR5 and CXCL13 and shed light on the origin and function of the MDV-encoded vIL-8 chemokine, which plays a crucial role in the pathogenesis of this highly oncogenic virus.