Karakterizacija podjedinice β (rpoB) RNA polimeraze vrste Ehrlichia canis izdvojene iz pasa i krpelja Rhipicephalus sanguineus u području Cebu na Filipinima.

Ehrlichia canis, a canine tick-borne pathogen with wide geographic distribution, has been serologically and molecularly detected in the Philippines. The present study aimed to characterize E. canis detected from Rhipicephalus sanguineus ticks and dogs in Cebu, Philippines, using the RNA polymerase sub-unit Beta (rpoB), a gene that has been used for disease diagnosis and resolution of phylogenetic relationships between closelyrelated species. Using a 16S rRNA gene-based PCR that screens Ehrlichia spp., DNA samples obtained from the blood of 10 dogs, confirmed to be serologically positive for E. canis, were tested and found positive for E. canis after subsequent DNA sequencing. DNA from infected ticks and the 16S rRNA-E. canis-positive canine blood samples from the present study were further analyzed using the rpoB gene. All registered Ehrlichia spp. rpoB gene sequences were aligned to design specific primers that can amplify a partial 1572-bp length sequence of E. canis. The obtained sequences revealed 99.8-100 % identities with each other, and 99.8-100 % and 87.8-89.1 % identities with registered E. canis and E. chaffeensis sequences from the USA, respectively. Phylogenetic analysis revealed that the obtained partial rpOB sequences formed a clade with E. canis strains from the USA. The present study is the first rpoB characterization of E. canis in the Philippines, and apparently in Asia, and provides additional evidence of the presence of the pathogen in the country. It also adds information on the high conservation of the rpoB gene in E. canis.Ehrlichia canis, geografski vrlo proširena rikecija u pasa, dokazana je serološki i molekularno na Filipinima. Određena su obilježja E. canis dokazane u krpelja Rhipicephalus sanguineus i pasa u pokrajini Cebu na Filipinima. To je učinjeno analizom podjedinice β (rpoB) RNA polimeraze, gena koji je rabljen za dijagnosticiranje bolesti i otkrivanje filogenetske srodnosti između usko srodnih vrsta. Upotrebom lančane reakcije polimerazom temeljene na genu 16S rRNA pomoću kojeg se razlikuju vrste roda Ehrlichia, u krvi 10 pasa serološki pozitivnih na E. canis dokazana je DNA specifična za E. canis. DNA iz zaraženih krpelja i uzorci krvi pasa pozitivnih na 16S rRNA-E. canis bili su dalje analizirani na osnovi gena rpoB. Sve dokazane sekvencije gena rpoB rikecija roda Ehrlichia bile su poravnate radi sinteze specifičnih početnica s kojima se može umnožiti slijed specifičan za E. canis dužine 1572 bp. Umnožene sekvencije pokazivale su 99,8-100 % identičnosti međusobno, 99,8-100 % identičnosti s vrstom E. canis iz SAD-a i 87,8-89,1 % identičnosti sa slijedovima E. chaffeensis iz SAD-a. Filogenetska analiza je pokazala da se sekvencije rpoB nalaze u skupini sa sojevima E. canis iz SAD-a. U istraživanju je prvi put analiziran rpoB vrste E. canis na Filipinima i u Aziji. Ono pruža dodatni dokaz prisutnosti te vrste na tom području. Također pruža informaciju o visokoj konzerviranosti gena rpoB vrste E. canis

Detection of gastrointestinal parasites in small-scale poultry layer farms in Leyte, Philippines

Background: Gastrointestinal (GIT) parasites can affect poultry productivity by compromising its health. It is well studied in other countries, but the documented reports in the Philippines have been limited. Aim: The aim of the present study was to evaluate the presence of GIT parasites in selected small-scale poultry layer farms in Leyte, Philippines. Materials and Methods: A total of 243 stool samples from eight small-scale poultry layer farms in Leyte, Philippines, were examined for GIT parasites using floatation and sedimentation technique. Profile parameters were also obtained. Fecal samples were collected and analyzed using floatation and sedimentation techniques. Statistical significance between GIT parasite positivity and profile parameters was determined using Chi-square test. Results: GIT parasites were detected in 92.2% of the samples (24.7% with single infection, 42.0% with 2-3 parasites, and 25.5% with three or more parasites). The common parasites detected were Ascaridia spp. (41.2%), Heterakis spp. (59.3%), Capillaria spp. (10.7%), Eimeria spp. (43.2%), and Strongyloides spp. (74.1%). Some profile parameters, including farm location, years in business, number of workers, nearby water system, the practice of fecal cleaning, and presence of other animals, were found to be significantly associated with GIT positivity. Conclusion: GIT parasites were detected in the poultry of small-scale layer farms in selected areas in Leyte, Philippines. This finding calls for the importance of routine GIT parasite monitoring and the implied need for regular deworming or dewormer rotation in the area

High Genetic Diversity of Anaplasma marginale Detected from Philippine Cattle

A total of 658 cattle in 6 provinces in the Philippines were screened for Anaplasma marginale infection by using a diagnostic heat-shock operon (groEL) gene-PCR assay. The screening-positive samples were further tested using the major surface antigen protein 1a (Msp1a) gene-PCR assay. Screening PCR results showed 130 cattle (19.8%) were positive for the A. marginale infection. Subsequent amplification using the Msp1a gene only showed 93 samples (14.1%) to be positive. In addition, 37 tandem-repeat structures, including 20 novel structures, and 41 distinct genotypes were identified. Interestingly, multiple infections of 4 different genotypes were also observed in A. marginale-infected cattle. The present study demonstrated the prevalence and characterization of diverse genotypes of A. marginale in the Philippine cattle.https://www.jstage.jst.go.jp/article/jvms/76/7/76_13-0405/_article/-char/ja

Endemicity of Toxoplasma infection and its associated risk factors in Cebu, Philippines.

Toxoplasma gondii is a single-celled intracellular apicomplexan parasite that causes toxoplasmosis. It is capable of infecting humans and nearly all warm-blooded animals including pigs, but cats are the only known definitive host. This ubiquitous zoonotic pathogen can cause abortion, stillbirth and fetal abnormalities, and has been associated with mental and behavioral changes in humans. Acute infection is potentially fatal in immunocompromised individuals. The present study aimed to assess the Toxoplasma seroprevalence in pigs, humans and cats after its initial reported detection in pigs about three decades ago in Cebu, Philippines. A total of 924 humans, 104 cats and 514 slaughter pigs were tested for antibodies against T. gondii using a commercial latex agglutination test. The results revealed positive detection rates of 26.3% (244/924) for humans, 42.3% (44/104) for cats and 13.4% (69/514) for slaughter pigs. Statistical analyses revealed that the area (P = 0.004), cat ownership (P = 0.020), the frequency of contact with cats (P 0.05). This study is the first report of the serological detection of T. gondii in humans and cats in Cebu, Philippines, and the first assessment of the prevalence of the parasite in pigs in the area since its initial detection in 1982. This is also the first report documenting the seropositivity of T. gondii in pregnant women in the country. The confirmed seropositivity of T. gondii in Cebu, Philippines, in the present study implies the endemicity of toxoplasmosis in this area and highlights the need for routine testing and increased public awareness

Retrospective analyses of dogs found serologically positive for Ehrlichia canis in Cebu, Philippines from 2003 to 2014

Aim: The study aimed to document the clinical and hematological observations of dogs found serologically positive for Ehrlichia canis and to identify parameters or factors that are associated with the disease with focus on the anemic and thrombocytopenic state of the infected dogs. Materials and Methods: From 7 participating veterinary establishments, a total of 913 cases from 2003 to 2014 were initially assessed using inclusion criteria, including E. canis diagnosis by the attending veterinarian and the presence of ticks or history of infestation, thrombocytopenia, and/or anemia. From these, 438 cases that were found serologically positive for E. canis using commercial test kits were selected. Profile, clinical observations and hematological test results were obtained from the selected cases. Computations for statistical associations between the anemic and thrombocytopenic state of the infected dogs and their profile, observed clinical signs and other hematological values were performed. Results: Most of the dogs were purebred (60.0%) and female (51.1%) and were within the age range of 1-5 years (38.4%). The mean packed cell volume (PCV), red blood cell (RBC) count, and platelet count were lower than the normal values while the absolute count of basophils were higher than normal values. Creatinine and blood urea nitrogen (BUN) appear to be elevated. The most common clinical signs observed were inappetence (41.3%), lethargy/depression (35.6%), vomiting (32.4%), fever (18.5%), paleness (8.2%), and epistaxis (6.6%). Analyses showed that there were no significant differences on the hematological values and clinical signs between thrombocytopenic and non-thrombocytopenic seropositive dogs. Moreover, very weak correlations between platelet count and RBC count, absolute lymphocyte count, and neutrophil count were found. On the other hand, only paleness (p=0.008) and epistaxis (p=0.004) were found to be significantly different between anemic and non-anemic patients. This coincided with the linear regression results where PCV (p=0.000, R=0.787, R2=0.619) was moderately correlated with the RBC count. In addition, eosinophil count was found weakly correlated. Conclusion: E. canis infection in dogs may produce varied clinical signs that may be influenced by the thrombocytopenic and anemic states of affected animals. Complete blood counts remain important in the diagnosis of the disease, especially the platelet and RBC counts. Creatinine, BUN and alanine aminotransferase can be of value in the diagnosis of the infection. Several cases were lost to follow-up and appeared to be a challenge for handling veterinarians to monitor compliance of owners and progress of infected patients

Profile and artificial insemination practices of technicians and the artificial insemination success rates in Leyte, Samar, and Biliran, Philippines (2011-2015)

Background: Artificial insemination (AI) is a reproductive biotechnology that may be influenced by several factors, including the profile of the technicians and the practices used. Assessing technician’s profile and their AI practices can be significant in improving AI success rate. Aim: This study aimed to know the profile and current practices used by AI technicians (AITs), to determine the success rates of AI in water buffaloes in Leyte, Samar, and Biliran from 2011 to 2015, and to evaluate the possible association between the parameters investigated. Materials and Methods: A total of 50 AITs from Leyte, Samar and Biliran, Philippines, were interviewed using a fixed questionnaire about their profile and employed AI practices, and 20,455 AI-related records of the Philippine Carabao Center (PCC) at Visayas State University (VSU), Baybay City, Leyte, were screened and analyzed. AI success rates were determined by retrospective analysis of the gathered data. Statistical analysis was performed between the technician profile and practices and the AI success rates. Results: Results revealed that most of the technicians were male, around 31-40 years old, married, college graduates, working under local government units, had other sources of income, and with 1-5 years of continuous AI practice averaging 51-100 inseminations per year. Most of them attended only one basic training seminar, which was conducted more than 3 years ago in PCC in VSU. AI success rates were recorded highest in 2011 and lowest in 2015. Statistical analyses showed that some technician profile parameters (civil status, average AI per year, and the training center) and several practices (checking of soft cervix, rectal palpation, thawing temperature method, straw cutting method, and semen deposition) might have an influence on the success of AI. Conclusion: This study documents the first report on AIT’s profile and their employed AI practices and the AI success rates in Leyte, Samar, and Biliran, Philippines. Selected profile parameters and AI practices may influence AI success rates. AITs should perform more AI services and revisit the employed practices

Evaluation on the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats (Capra hircus) in Cebu, the Philippines

Background: Tick-borne diseases are caused by a wide variety of viruses, pathogens, and diseases. Anaplasma, Ehrlichia, and Babesia spp. are among the most known tick-borne pathogens in Asia. In the Philippines, these pathogens were already reportedly present in dogs and large ruminants, but no study has been reported yet evaluating their presence in goats. Aim: The present study aimed to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. in goats in Cebu, the Philippines. Materials and Methods: A total of 100 blood samples from goats were collected in Cebu, the Philippines. Profile of sampled goats including age, body score, and sex was obtained. Peripheral blood smear examination and DNA extraction were performed. Nested polymerase chain reaction assay was used to evaluate the presence of Anaplasma, Ehrlichia, and Babesia spp. Results: None of the samples were found positive with Anaplasma, Ehrlichia, and Babesia spp. infection. Conclusion: Tested goats were negative with Anaplasma, Ehrlichia, and Babesia spp. and calls for continuous surveillance of these pathogens due to the reported detection of these pathogens in other livestock animals in the area