Hala El Daous

Benha University

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Co-Authors

Shuya Mitoma

University of Miyazaki

Satoshi Sekiguchi

University of Miyazaki

Junzo Norimine

University of Miyazaki

Kosuke Notsu

University of Miyazaki

Thi Ngan

Vietnam National University of Agriculture

Eslam Elhanafy

Benha University

Emmanuel Kabali

Food and Agriculture Organization of the United Nations

Genki Arikawa

Hokkaido University

Xinyue Wu

Liaoning University of Traditional Chinese Medicine

Heba Elkhaiat

Benha University

Cooperative Institutions

University of Miyazaki

Benha University

Hokkaido University

University of Zambia

Ministry of Agriculture and Livestock

Agricultural Research Center

The University of Tokyo

Jeonbuk National University

Zagazig University

University of Edinburgh

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Identifying Pathogen and Allele Type Simultaneously (IPATS) in a single well using droplet digital PCR

bioRxiv (Cold Spring Harbor Laboratory) (2022)

Kosuke Notsu Hala El Daous Shuya Mitoma Xinyue Wu Junzo Norimine

Abstract A combined host biomarker and pathogen diagnosis provides insight into disease progression risk and contributes to appropriate clinical decision-making regarding prevention and treatment. In preventive veterinary medicine, such combined diagnosis could improve risk-based livestock herd management. We developed a single-well based test for combined diagnosis of bovine leukemia virus (BLV) and bovine MHC ( BoLA )- DRB3 alleles. A fourplex droplet digital PCR method targeting the BLV pol gene, BLV-susceptible DRB3*016:01 allele, resistant DRB3*009:02 allele, and housekeeping RPP30 gene (IPATS-BLV) successfully measured the percentage of BLV-infected cells and determined allele types precisely. Furthermore, it discriminated homozygous from heterozygous carriers. Using this method to determine the impact of carrying these alleles on the BLV proviral load (PVL), we found DRB3*009:02-carrying cattle could suppress the PVL to a low or undetectable level, even with the presence of a susceptible allele. Although the population of DRB3*016:01 -carrying cattle showed significantly higher PVLs when compared with cattle carrying other alleles, their individual PVLs were highly variable. Because of the simplicity and speed of this single-well assay, IPATS could be a suitable platform for the combined diagnosis of host biomarkers and pathogens in a wide range of other systems.

Bovine leukemia virus

10.1101/2022.09.10.507438

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Quantitative Risk Assessment for the Introduction of Bovine Leukemia Virus-Infected Cattle Using a Cattle Movement Network Analysis

Pathogens (2020)

Kosuke Notsu Anuwat Wiratsudakul Shuya Mitoma Hala El Daous Chiho Kaneko

The cattle industry is suffering economic losses caused by bovine leukemia virus (BLV) and enzootic bovine leukosis (EBL), the clinical condition associated with BLV infection. This pathogen spreads easily without detection by farmers and veterinarians due to the lack of obvious clinical signs. Cattle movement strongly contributes to the inter-farm transmission of BLV. This study quantified the farm-level risk of BLV introduction using a cattle movement analysis. A generalized linear mixed model predicting the proportion of BLV-infected cattle was constructed based on weighted in-degree centrality. Our results suggest a positive association between weighted in-degree centrality and the estimated number of introduced BLV-infected cattle. Remarkably, the introduction of approximately six cattle allowed at least one BLV-infected animal to be added to the farm in the worst-case scenario. These data suggest a high risk of BLV infection on farms with a high number of cattle being introduced. Our findings indicate the need to strengthen BLV control strategies, especially along the chain of cattle movement.

Bovine leukemia virus

Enzootic

Cattle Diseases

Holstein Cattle

10.3390/pathogens9110903

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Citations (9)

Efficacy of dietary supplements of Glycyrrhiza glabra (Licorice) and maduramicin alone or in combination with Eimeria tenella infected chicks: A clinical study and molecular docking

Open Veterinary Journal (2024)

Sawsan S. Elbasuni Hanan A. A. Taie Samah M. Abdel Gawad Reda E L Kamar Hala El Daous

Background: Coccidiosis is one of the most economically significant poultry diseases worldwide, caused by the pathogenic Eimeria species, and is characterized by decreased weight gain and failure to grow due to malabsorption, low feed conversion rate, bloody diarrhea, and dehydration. Aim: This study investigated the effectiveness of licorice root extract (LRE) in controlling cecal coccidiosis to determine whether its combination with maduramicin could help alleviate the pathological, biochemical, and histopathological effects of cecal coccidiosis in Sasso broiler chicks. Methods: A total of 125 one-day-old Sasso broiler chicks were categorized into five equal groups (n=25), each consisting of five replicates (n= 5 per replicate). (G1-LE) received a basal diet supplemented with LRE (3 g/kg); (G2-ME) received a basal diet containing maduramycin (0.5 g/kg); (G3-LME) received a basal diet containing LRE and maduramicin together with the same rates. G4-E (positive control) and G5-N (negative control) received no additives in their feed. Birds in groups (G1-4) were challenged on day 14 of the experiment orally intercropping a 1 ml suspension of Eimeria tenella sporulated oocysts. Results: Groups of birds fed on LRE and maduramicin separately or together appeared to be in good condition where no deaths or clinical abnormalities were observed, based on the analysis of clinicopathological examination. Compared with the G4-E positive control, the dropping scoring and oocyst shedding of groups G1-LE, G2-ME, and G3-LME along the 10th-day post-challenge (dpc), as well as macroscopic and microscopic lesions scoring at the 7th dpc, was considerably lower. The dual supplementation use of LRE and maduramicin in G3-LME's reduced the harmful effects of coccidian, which appeared only as a mononuclear cellular infiltration and a small number of oocysts invading the intestinal glands. Molecular docking revealed that LRE and maduramicin interacted with E. tenella DNA polymerase, E. tenella apical membrane antigen 1, and microneme protein binding sites resulting in reduced E. tenella replication and invasion. Conclusion: The inclusion of LRE and maduramicin, individually or in combination, in the diet might effectively mitigate the detrimental effects of coccidiosis.

10.5455/ovj.2024.v14.i1.20

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Evaluating the Risk Factors for Porcine Epidemic Diarrhea Virus Infection in an Endemic Area of Vietnam

Frontiers in Veterinary Science (2020)

Thi Ngan Thanh Phong Bui Thi My Le Huynh Yosuke Sasaki Shuya Mitoma

Porcine epidemic diarrhea virus (PEDV) causes enteritis, vomiting, watery diarrhea, and high mortality in suckling pigs, threatening the swine industry. Porcine epidemic diarrhea (PED) re-emerged globally in 2013 in many important swine-producing countries in Asia and the Americas. Several studies have identified the risk factors for the spread of PEDV in acute outbreaks. However, limited information is available on the risk factors for the transmission of PEDV in endemic regions. We hypothesized that poor biosecurity, location, and some social or cultural practices are the main risk factors for PEDV transmission in the Vietnamese pig population. The aim of this study was to evaluate the potential risk factors for the transmission of PEDV in an endemic area in Vietnam. In this case-control study, questionnaires containing 51 questions were completed for 92 PEDV-positive and 95 PEDV-negative farms. A logistic regression analysis was performed to assess the risk factors associated with PEDV infection. Province and the total number of pigs were included as random effects to determine their influence on the risk of PEDV infection. Twenty-nine variables of interest that have been associated with PEDV status were analyzed in a univariate analysis (P <0.20), with backward stepwise selection. Only three of these 29 variables in four models remained significant PEDV risk factors in the final model: farrow-to-wean production type, distance from the farm to the slaughterhouse (<1,000 m), and the presence of chickens on site (P <0.05). This is the first study to identify the main risk factors for PEDV infection in an endemic area. Our findings suggest that hygiene measures should be strictly implemented on farms for the effective control and prevention of PEDV infection.

Biosecurity

Vietnamese

10.3389/fvets.2020.00433

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Slaughterhouse survey for detection of bovine viral diarrhea infection among beef cattle in Kyushu, Japan

Journal of Veterinary Medical Science (2019)

Mohammad Aref AGAH Kosuke Notsu Heba Elkhaiat Genki Arikawa Meiko Kubo

Bovine viral diarrhea virus (BVDV) footprint has spread across the globe and is responsible for one of the most economically important diseases in cattle. In Japan, some regional surveillance and preventive measures to control bovine viral diarrhea (BVD) have been implemented. However, BVDV infection is poorly understood in cattle industries, and there is no systematic BVD surveillance system and control program. Kyushu is the center for raising beef cattle in Japan. Therefore, this study aimed to determine the BVDV infection using a slaughterhouse survey among beef cattle in Kyushu, Japan. A total of 1,075 blood samples were collected at two regional slaughterhouses in Miyazaki prefecture from December 2015 to June 2016. Antigen ELISA was used for detection of BVDV antigen in blood samples. Two samples showed positive results (2/1,075; 0.18%). BVDV RNA was extracted from positive blood samples; the sequence was determined and analyzed by the neighbor-joining method for construction of the phylogenetic tree. Phylogenetic analysis based on the 5'-UTR revealed that the two positive samples were grouped into the same subtype BVDV-1b in the BVDV-1 genotype, but the infected cattle belonged to two different farms. In conclusion, this is the first study to identify the presence of BVDV in a slaughterhouse survey in Kyushu. These findings suggest that a slaughterhouse survey is a useful tool for developing a surveillance system for monitoring infectious diseases in cattle.

Beef Cattle

Cattle Diseases

10.1292/jvms.19-0045

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Citations (3)

Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA

Journal of Virological Methods (2023)

Xinyue Wu Kosuke Notsu Yuichi Matsuura Shuya Mitoma Hala El Daous

Bovine leukemia virus

Provirus

genomic DNA

TaqMan

10.1016/j.jviromet.2023.114706

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Citations (7)

Corrigendum to “Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA” [J. Virol. Methods, 315 (2023) 114706]

Journal of Virological Methods (2023)

Xinyue Wu Kosuke Notsu Yuichi Matsuura Shuya Mitoma Hala El Daous

Bovine leukemia virus

genomic DNA

10.1016/j.jviromet.2023.114708

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A pooled testing system to rapidly identify cattle carrying the elite controllerBoLA‐DRB3*009:02haplotype against bovine leukemia virus infection

HLA (2021)

Kosuke Notsu Hala El Daous Shuya Mitoma Junzo Norimine Satoshi Sekiguchi

As genetically resistant individuals, the "elite controllers" (ECs) of human immunodeficiency virus infection have been focused on as the keys to developing further functional treatments in medicine. In the livestock production field, identifying the ECs of bovine leukemia virus (BLV) infection in cattle is desired to stop BLV transmission chains on farms. Cattle carrying the bovine leukocyte antigen (BoLA)-DRB3*009:02 allele (DRB3*009:02) have a strong possibility of being BLV ECs. Most of cattle carrying this allele maintain undetectable BLV proviral loads and do not shed virus even when infected. BLV ECs can act as transmission barriers when placed between uninfected and infected cattle in a barn. To identify cattle carrying DRB3*009:02 in large populations more easily, we developed a pooled testing system. It employs a highly sensitive, specific real-time PCR assay and TaqMan MGB probes (DRB3*009:02-TaqMan assay). Using this system, we determined the percentage of DRB3*009:02-carrying cattle on Kyushu Island, Japan. Our pooled testing system detected cattle carrying the DRB3*009:02 allele from a DNA pool containing one DRB3*009:02-positive animal and 29 cattle with other alleles. Its capacity is sufficient for herd-level screening for DRB3*009:02-carrying cattle. The DRB3*009:02-TaqMan assay showed high-discriminative sensitivity and specificity toward DRB3*009:02, making it suitable for identifying DRB3*009:02-carrying cattle in post-screening tests on individuals. We determined that the percentage of DRB3*009:02-carrying cattle in Kyushu Island was 10.56%. With its ease of use and reliable detection, this new method strengthens the laboratory typing for DRB3*009:02-carrying cattle. Thus, our findings support the use of BLV ECs in the field.

Bovine leukemia virus

TaqMan

10.1111/tan.14502

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Citations (6)

Assessment of Hematological Parameters and Carcass Weight in Bovine Leukemia Virus Infection in Slaughtered Beef Cattle

Journal of Veterinary Epidemiology (2018)

Kosuke Notsu Shiori Hashida Shuya Mitoma Meiko Kubo Genki Arikawa

Most bovine leukemia virus (BLV)-infected cattle do not have clinical signs (aleukemic AL), but some develop persistent lymphocytosis (PL) and B-cell lymphosarcoma (enzootic bovine leucosis [EBL]). BLV infection is a well-known cause of chronic wasting disease, which is associated with a reduction in milk productivity and immunity in dairy cattle. However, the effect of BLV infection on beef cattle is not clear. The objective of this study was to investigate the effect of BLV infection on the productivity of slaughtered beef cattle. A total of 997 blood samples were collected from cattle in 2 slaughterhouses in Miyazaki prefecture, Japan. BLV-antibodies were tested in these cattle's blood samples using enzyme-linked immunosorbent assay (ELISA), to identify BLV-infected cattle. We compared blood parameters and carcass weight between BLV ELISA-positive and ELISA-negative cattle in two age groups : young (≤ 60 months) and elder (> 60 months) groups. The results showed that the proportion of ELISA-positive cattle in the young and elder groups were 22.8% and 24.9%, respectively. The number of white blood cells (WBCs) and lymphocytes in ELISA-positive cattle was significantly higher than that in ELISA-negative cattle in the young group. In addition to the number of lymphocytes, the number of monocytes and neutrophils were also significantly higher in BLV ELISA-positive cattle than in ELISA-negative cattle in the elder group. There was no significant difference in the carcass weight between ELISA-positive and ELISA-negative cattle in both groups. The results of this study suggest that BLV infection has an effect on the host immune response in beef cattle.

Bovine leukemia virus

Leucosis

Enzootic

Beef Cattle

White blood cell

Lymphocytosis

10.2743/jve.22.43

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Citations (2)

Assessment of biochemical biomarkers and environmental stress indicators in some freshwater fish

Environmental Geochemistry and Health (2024)

Salwa M. Abdallah Reham E. Muhammed Reda El Mohamed Hala El Daous Dina Saleh