Using geographic rescue time contours, point-of-care strategies, and spatial care paths to prepare island communities for global warming, rising oceans, and weather disasters.

OBJECTIVES: To perform geographic contour analysis of sea and land ambulance rescue times in an archipelago subject to super typhoons; to design point-of-care testing strategies for medical emergencies and weather disasters made more intense by global warming and rising oceans; and to assess needs for prehospital testing on spatial care paths that accelerate decision making, increase efficiency, improve outcomes, and enhance standards of care in island nations. METHODS: We performed needs assessments, inspected healthcare facilities, and collected ambulance rescue times from professionals in the Bantayan Archipelago, Philippines. We mapped sea/land ambulance rescue routes and time contours. To reveal gaps, we statistically compared the fastest and slowest patient rescue times from islands/islets and barangays to the District Hospital on Bantayan Island. We developed spatial care paths (the fastest routes to care) for acute myocardial infarction, community care, and infectious diseases. We generated a compendium of prehospital diagnostic testing and integrated outcomes evidence, diagnostic needs, and public health goals to recommend point-of-care strategies that build geographic health resilience. RESULTS: We observed limited access to COVID-19 assays, absence of blood gas/pH testing for critical care support, and spatial gaps in land and airborne rescues that worsened during inclement weather and sea swells. Mean paired differences (slowest-fastest) in ambulance rescue times to the District Hospital for both islands and barangays were significant (P < 0.0001). Spatial care path analysis showed where point-of-care cardiac troponin testing should be implemented for expedited care of acute myocardial infarction. Geospatial strengths comprised distributed primary care that can be facilitated by point-of-care testing, logical interisland transfers for which decision making and triage could be accelerated with onboard diagnostics, and healthcare networks amenable to medical advances in prehospital testing that accelerate treatment. CONCLUSIONS: Point-of-care testing should be positioned upstream close to homes and island populations that have prolonged rescue time contours. Geospatially optimized point-of-need diagnostics and distributed prehospital testing have high potential to improve outcomes. These improvements will potentially decrease disparities in mortality among archipelago versus urban dwellers, help improve island public health, and enhance resilience for increasingly adverse and frequent climate change weather disasters that impact vulnerable coastal areas. [350 words].

Bovine Piroplasma Populations in the Philippines Characterized Using Targeted Amplicon Deep Sequencing.

Molecular assays and capillary electrophoresis sequencing have been used to identify parasites in livestock. The low sample capacity, which increases labor and processing time, is one drawback. Targeted amplicon sequencing (Ampliseq) uses the fast and large sample capacity platform to identify parasites in the target host, overcoming this limitation. DNA was extracted from 162 whole blood samples collected from cattle in three provinces in the Philippines. Using Illumina's Miseq platform, the V4 hypervariable region of the piroplasma 18S rRNA gene was amplified and sequenced. The AMPtk pipeline was used to obtain distinct amplicon sequence variants (ASVs) and the NCBI BLAST non-redundant database was used to assign taxonomy. In total, 95 (58.64%) samples were positive for piroplasma. Using the AMPTk pipeline, 2179 ASVs were obtained. A total of 79 distinct ASVs were obtained after clustering and filtering, which belonged to genera Babesia (n = 58), Theileria (n = 17), Hepatozoon (n = 2), and Sarcocystis (n = 2). The ASV top hits were composed of 10 species: Babesia bovis, B. bigemina, Theileria orientalis, Babesia sp., Hepatozoon canis, Sarcocystis cruzi, T. annulata, T. equi, T. mutans, and Theileria sp. Thung Song. The results generated in this study demonstrated the applicability of Ampliseq in detecting piroplasmid parasites infecting cattle in the Philippines.

General knowledge and practice of household dog owners regarding gastrointestinal parasites in Cebu, Philippines.

Background and Aim: Canine gastrointestinal tract (GIT) parasites are of public health and veterinary concern and are harmful to both humans and animals. The risk of transmitting GIT parasites can be minimized depending on dog owners' knowledge and management practices. Therefore, this study aimed to assess dog owners' general knowledge and practices regarding GIT parasites. Materials and Methods: A structured questionnaire containing 10-point Likert statements was administered to 130 respondents in Cebu, Philippines, to obtain information about their profile (age, sex, civil status, and educational attainment) and data regarding their home environment, number of dogs and other pets owned, and their knowledge and practices regarding canine GIT parasites. These respondents had previously provided canine stool samples for fecalysis. Results: Most respondents were female (65.4%), 18-24 years old (55.4%), single (71.5%), and educated to college level (49.2%). Housing styles were mostly gated (76.9%), and >50% had a garden. The majority owned 1-3 dogs (56.9%) and did not own any other pets (73.8%). All knew about canine GIT parasites. There was a significant association between GIT parasite positivity in dogs and the owner's sex (p = 0.005). Gastrointestinal parasite positivity was also associated with the owner's knowledge about the transmission of parasites from the mother's milk to her puppies (p = 0.001), canine intestinal parasites potentially causing weight loss, diarrhea, and vomiting (p = 0.001), and dogs potentially becoming infected with parasites from licking or sniffing the soil or grass where other animals have been (p = 0.005). Moreover, there was a significant association between GIT parasite positivity and the owner's practice of daily cleaning of the areas where the dog defecated (p = 0.001), deworming every 6-12 months (p = 0.001), and unfailingly following the vaccination and deworming schedule (p = 0.006). Finally, the summative knowledge and practice level of the owners were significantly associated with GIT parasite positivity (p = 0.001). Conclusion: This study highlights the need for continuous education of dog owners to maintain good knowledge and practices to prevent GIT parasite infection. Frequent deworming (once a month) of adult dogs is recommended.

Molecular Detection of Animal Trypanosomes in Different Animal Species in the Visayas Region of the Philippines.

PURPOSE: Animal trypanosomosis is one of the most important parasitic diseases significantly affecting the Philippine economy. It is considered by the government to be the second most important disease of livestock after fasciolosis. A PCR-based molecular survey for trypanosomes in different animals in Bohol, Philippines, was performed to assess the prevalence of trypanosomosis in the area during the rainy and dry season. METHODS: A total of 269 blood samples were collected in two batches in rainy and dry season from different animal species in Ubay Stock Farm in Ubay, Bohol, the Philippines, including 151 samples from water buffaloes, 76 samples from cattle, 35 samples from goats, and 7 samples from horses. DNA was subsequently extracted from these blood samples, and two different PCR assays were employed to detect and identify trypanosomes DNA including ITS1 PCR and CatL PCR. RESULTS: Animal trypanosomes, Trypanosoma evansi and Trypanosoma theileri, were detected in water buffalo (37.7%) [95%CI: 30.4 - 45.7], cattle (44.7%) [95%CI: 34.1 - 55.9], and goats (34.3%) [95%CI: 20.8 - 50.8]. Only T. evansi was detected in horses (28.6%) [95% CI: 8.2 - 64.1]. No clinical signs were observed in all positive animals. CONCLUSION: This highlights the importance of domestic animals that can be infected with no signs but may act as reservoir animals and transmit trypanosomosis to susceptible animals. This study supports the importance of regular surveillance to estimate the prevalence of the disease, emphasizing its various dynamics in the affected areas and supporting efficient intervention measures.

Heartworm adulticide treatment: a tropical perspective.

Dirofilaria immitis (the canine heartworm) is widespread in the tropics, with prevalence surpassing 30% in high-risk areas. In addition to the suitable climatic conditions that favour mosquito abundance and filarial larva development, there is low compliance with the recommended year-round use of preventives in these transmission hotspots. This represents a major concern, considering that melarsomine (first-line heartworm adulticide) is unavailable in several tropical countries, resulting in the so-called slow-kill protocol being the only available adulticide treatment option. In this article, the members of TroCCAP (Tropical Council for Companion Animal Parasites) review the current distribution of heartworm in the tropics and the availability of melarsomine, and discuss alternatives for the management of heartworm infections in dogs.

First Molecular Identification of Babesia, Theileria, and Anaplasma in Goats from the Philippines.

Goats are key livestock animals and goat raising is an income-generating venture for smallholder farmers, supporting agricultural development in many parts of the world. However, goat production is often limited by various factors, such as tick-borne diseases. Goat piroplasmosis is a disease caused by apicomplexan parasites Babesia spp. and Theileria spp., while anaplasmosis is caused by bacterial Anaplasma spp. In the Philippines, the presence of Babesia, Theileria, and Anaplasma has not been reported in goats. In this study, DNA obtained from goats were molecularly screened for Babesia/Theileria and Anaplasma. Of 396, 77.02% (305/396) and 38.64% (153/396) were positive for piroplasma and Anaplasma using PCR assays targeting the 18S rRNA and 16S rRNA genes, respectively. Similarly, Babesia ovis was detected in six samples (1.52%). Representative Babesia/Theileria sequences shared 89.97-97.74% identity with each other and were most closely related to T. orientalis, T. annulata, and Theileria spp. Meanwhile, Anaplasma 16SrRNA sequences were related to A. odocoilei, A. platys, and A. phagocytophilum. This is the first molecular identification of B. ovis, Theileria spp., and Anaplasma spp. in goats from the Philippines.

Molecular Identification of Selected Tick-Borne Protozoan and Bacterial Pathogens in Thoroughbred Racehorses in Cavite, Philippines.

Tick-borne diseases (TBDs) considerably impair equine health and productivity. Moreover, TBDs, particularly equine piroplasmosis, impede international movement and trade of equids, which is a vital component of the global horse racing industry. In the Philippines, horse racing is a lucrative industry generating millions of USD annually. However, information on equine TBDs is scarce. This study intended to describe molecularly the equine tick-borne infections in a racehorse park in Cavite, Philippines and identify the risk factors associated with the infections. One hundred twenty-four (n = 124) thoroughbred racehorses were sampled and screened for selected tick-borne protozoan and bacterial pathogens using polymerase chain reaction (PCR) assays. Racehorses were positive for Babesia caballi (12.10%; 15/124), Theileria equi (0.81%; 1/124), Anaplasma phagocytophilum (10.48%; 13/124), Borrelia burgdorferi sensu lato (38.71%; 48/124), A. marginale (0.81%; 1/124), and Coxiella burnetii (0.81%; 1/124). Rickettsia was not detected in the samples. Gender was determined as a significant risk factor for B. caballi infection. Sequencing analysis revealed that seven partial 18S rRNA B. caballi isolates shared 98.63-100% identity with each other and were classified as genotype A. Meanwhile, the sequence obtained from the lone T. equi-positive sample was 99.77% identical to isolates from Spain, Switzerland, China, Saudi Arabia, and South Korea, and was confirmed as genotype E based on the 18S rRNA gene. Eight Anaplasma 16S rRNA partial sequences were highly identical to A. phagocytophilum and A. ovis. Partial sequences of Borrelia 5-23S rRNA were most closely related to B. japonica and other Borrelia sp. isolates from various countries. This study reports the first molecular detection of Borrelia and Anaplasma and the identification of B. caballi and T. equi genotypes in racehorses in the Philippines. Findings from this study shall be useful in crafting equine tick and TBD control and prevention programs in the country.

First molecular detection and identification of Trypanosoma evansi in goats from Cebu, Philippines using a PCR-based assay.

The increasing number and severity of surra outbreaks in the Philippines led the government to consider it as the second most important disease of livestock in the country. It is one of the most economically important animal parasitic diseases and has been reported in several animal species, including water buffaloes, cattle, and horses in different regions of the Philippines. However, it has not yet been reported in Cebu, the usual gateway of livestock trade in the area that raises 6% of the 3.75 million goats in the country. In the current study, a PCR-based assay was conducted for the molecular detection and characterization of Trypanosoma evansi in goats in Cebu. A total of 251 goats were randomly sampled from four farms. DNA was extracted and ITS1-PCR was applied to detect different trypanosomes in goats. Eighty-five out of the 251 (33.9%) samples tested positive for T. evansi, two of which were also positive for T. theileri-like trypanosome. The detection rate of T. evansi was slightly higher in male goats (38.3%) than in females (32.5%), and in younger goats (34.5%) than in adults (33.5%). The findings, however, did not differ significantly to suggest any association between sex and age with T. evansi infection in goats. The detection of T. evansi and T. theileri-like trypanosome in goats was confirmed by sequence analysis of ITS1 region. To our knowledge, this is the first report on the molecular detection and identification of caprine T. evansi infection in Cebu, Philippines.

TroCCAP recommendations for the diagnosis, prevention and treatment of parasitic infections in dogs and cats in the tropics.

The Tropical Council for Companion Animal Parasites Ltd. (TroCCAP) is a not-for-profit organisation whose mission is to independently inform, guide and make best-practice recommendations for the diagnosis, treatment and control of companion animal parasites in the tropics and sub-tropics, with the aim of protecting animal and human health. In line with this primary mission, TroCCAP recently developed guidelines for the diagnosis, treatment and control of feline and canine parasites in the tropics. The development of these guidelines required unique and complex considerations to be addressed, often inapplicable to developed nations. Much of the tropics encompass middle-to-low income countries in which poor standards of environmental hygiene and large populations of stray dogs and cats coexist. In these regions, a range of parasites pose a high risk to companion animals, which ultimately may place their owners at risk of acquiring parasitic zoonoses. These considerations led to the development of unique recommendations with regard, for example, to deworming and endoparasite testing intervals for the control of both global and 'region-specific' parasites in the tropics. Moreover, the 'off-' or 'extra'-label use of drugs for the treatment and control of parasitic infections is common practice in many tropical countries and many generic products lack manufacturers' information on efficacy, safety, and quality control. Recommendations and advice concerning the use of such drugs and protocols are also addressed in these guidelines. The formation of these guidelines is an important first step towards improving the education of veterinarians specifically regarding best-practice for the diagnosis, treatment and control of canine and feline parasites in the tropics.

Molecular survey of tick-borne pathogens infecting backyard cattle and water buffaloes in Quezon province, Philippines.

Tick-borne diseases (TBD) cause enormous losses for farmers. Backyard raising comprises majority of the livestock population in the Philippines, but TBD information in backyard livestock is scarce. In this study, 48 cattle and 114 water buffalo samples from Quezon province, Philippines were molecularly screened for tick-borne pathogens. Anaplasma marginale (16.67%) and hemoplasma (20.99%) were detected in the samples. A. marginale infection (P=0.0001) was significantly higher in cattle, while hemoplasma infection (P=0.011) was significantly higher in water buffaloes. A. marginale isolates from this study were highly similar to previous isolates from the Philippines while Mycoplasma wenyonii and Candidatus Mycoplasma haemobos were the identified hemoplasma species. Our findings reveal additional information on the TBD situation of Philippine backyard livestock.

Review on the Current Trends of Toxoplasmosis Serodiagnosis in Humans.

Toxoplasmosis is a widely distributed zoonotic infection caused by the obligate intracellular apicomplexan parasite Toxoplasma gondii. It is mainly transmitted through the ingestion of oocysts shed by an infected cat acting as its definitive host. The key to effective control and treatment of toxoplasmosis is prompt and accurate detection of T. gondii infection. Several laboratory diagnostic methods have been established, including the most commonly used serological assays such as the dye test (DT), direct or modified agglutination test (DAT/MAT), indirect hemagglutination test (IHA), latex agglutination test (LAT), indirect immunofluorescent test (IFAT), enzyme-linked immunosorbent assays (ELISA), immunochromatographic tests (ICT), and the western blot. Nonetheless, creating specific and reliable approaches for serodiagnosis of T. gondii infection, and differentiating between acute and chronic phases of infection remains a challenge. This review provides information on the current trends in the serodiagnosis of human toxoplasmosis. It highlights the advantages of the use of recombinant proteins for serological testing and provides insight into the possible future direction of these methods.

Host range and geographical distribution of Babesia sp. Mymensingh.

Bovine babesiosis represents a serious threat to the cattle industry in the tropics and subtropics. Although several Babesia species infect cattle, only B. bovis, B. bigemina and B. divergens are known to cause clinical babesiosis. However, our recent study demonstrated that the newly discovered Babesia sp. Mymensingh might be a virulent species capable of causing clinical babesiosis in cattle. The objective of this study was to determine the host range and geographical distribution of Babesia sp. Mymensingh on a global scale. A total of 2,860 archived DNA samples from 2,263 cattle in Sri Lanka (n = 672), the Philippines (n = 408), Vietnam (n = 460), Uganda (n = 409), Brazil (n = 164) and Argentina (n = 150); 419 buffalo in Sri Lanka (n = 327) and Vietnam (n = 92); and 127 goats and 51 sheep in Vietnam were screened using a Babesia sp. Mymensingh-specific PCR assay. Babesia sp. Mymensingh infection was detected in cattle, buffalo, sheep and goats. Cattle of all countries surveyed in this study except Brazil were found to be infected with Babesia sp. Mymensingh. The highest positive rates were recorded in cattle from the Philippines (11.3%) and Vietnam (9.6%), followed by Argentina (4.7%), Sri Lanka (1.5%) and Uganda (1.0%). Buffalo were found to be infected with this parasite in Sri Lanka (1.2%) and Vietnam (10.9%). Unexpectedly, Babesia sp. Mymensingh was also detected in sheep (2.0%) and goats (1.3%) from Vietnam. These findings were confirmed by PCR amplicon sequencing. In conclusion, our present findings indicate that Babesia sp. Mymensingh, which infects cattle, buffalo, sheep and goats, is endemic in Asia, Africa and South America.

Detection of canine Schistosoma japonicum infection using recombinant thioredoxin peroxidase-1 and tandem repeat proteins.

Humans and dogs live very close together and share various pathogens causing zoonotic parasitoses like schistosomiasis. A previous population genetics study done for schistosomes in the Philippines suggested that there is a high transmission level of Schistosoma japonicum among humans and dogs proving that the latter are important reservoirs for this zoonotic parasite. A more sensitive and specific test detecting schistosome infection in dogs will therefore strengthen the zoonotic surveillance, which might help in the possible elimination of this ancient disease. In this study, recombinant thioredoxin peroxidase-1 (SjTPx-1) and tandem repeat proteins (Sj1TR, Sj2TR, Sj4TR, Sj7TR) previously tested on human and water buffalo samples were used to assess its diagnostic applicability to dogs. Fifty-nine dog serum and stool samples were collected in the schistosomiasis-endemic municipalities of Calatrava, Negros Occidental and Catarman, Northern Samar in the Philippines and examined using the ELISA as compared to microscopy and fecal sample-based PCR. Samples positive for Babesia gibsoni and Dirofilaria immitis were also used to check for cross-reaction. Results showed that SjTPx-1 (80% sensitivity, 92.3% specificity) and Sj7TR (73.3% sensitivity, 92.3% specificity) have good potentials for diagnosing S. japonicum infection in dogs. These diagnostic antigens will therefore improve the surveillance in the transmission of the parasites from dogs to humans.

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.

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 < 0.0001) and consumption of street foods (P = 0.043) were significantly associated with seropositivity for T. gondii in humans. Meanwhile, the use of litter trays (P = 0.001) and contact with other animals (P = 0.007) were significantly associated with seropositivity in cats. The odds ratio for selected significant factors revealed that living in suburban areas (OR 1.66, 95% CI: 1.20-2.31), owning a cat (OR 1.482, 95% CI: 1.07-2.07) and eating street foods (OR 1.585, 95% CI: 1.01-2.48) were associated with an increased risk of T. gondii exposure in humans. In cats, the use of a litter tray (OR 4.5, 95% CI: 1.73-11.71) was associated with an increased risk of exposure. None of the profile parameters were found to be significantly associated with seropositivity in slaughter pigs (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.

The detection of gastrointestinal parasites in owned and shelter dogs in Cebu, Philippines.

BACKGROUND: Gastrointestinal tract (GIT) parasites affect the health of dogs and may also be zoonotic. The prevalence of these parasites has been well studied in several countries, but reports in the Philippines have been limited. AIM: This study generally aimed to detect the presence of common GIT parasites in owned and shelter dogs in Cebu, Philippines. MATERIALS AND METHODS: A total of 200 fecal samples (130 from owned dogs and 70 from shelter dogs) were collected. Profiles of owned dogs and their owners were obtained. Fecalysis was performed using three methods: Direct smear, sedimentation, and flotation techniques. RESULTS: Majority of the sampled dogs were 5 years old and below that (79.2%), male (64.6%) and of pure breed (53.1%). Among the most common parasites detected were Ancylostoma, Trichuris and Toxocara spp. Statistical analyses revealed a significant association between the presence of parasites and the body score of the dogs (p=0.000), the deworming status (p=0.000), and the rearing practice (contact with other dogs, p=0.000, where it spends its time (p=0.000), plays in the grass (p=0.050), where it defecates (p=0.014), contact with other animals (p=0.000). CONCLUSION: GIT parasites were detected in owned and shelter dogs in Cebu, Philippines. The results of this study can serve as baseline information about the canine parasitic fauna in the Philippines.

Molecular evidence of hemotropic mycoplasmas in goats from Cebu, Philippines.

Hemotropic mycoplasma (hemoplasma), a neglected vector-borne pathogen in goats, causes extensive economic damage to farmers due to production losses. In this study, 107/295 (36.27%) goats sampled from 4 farms (Barili, Danao City, Dumanjug and Minglanilla) in Cebu, Philippines tested positive for PCR targeting the 16S rRNA gene of Mycoplasma. All hemoplasma-positive goats were from Barili and no clinical sign was observed. Sex (P=0.0005) and age (P=0.03) were found associated with hemoplasma infection. Mycoplasma ovis, Candidatus Mycoplasma haemobos, Candidatus Mycoplasma haemominutum and 3 Uncultured Mycoplasma sp. sequences were identified by sequencing analysis. This is the first report of molecular detection and genetic characterization of hemoplasmas in goats in the Philippines.

First molecular detection and characterization of tick-borne pathogens in water buffaloes in Bohol, Philippines.

The water buffalo industry is a vital part of the Philippine livestock economy and is an essential contributor to the developing local dairy industry. Although relatively less susceptible to diseases, water buffaloes can still be infected and can act as reservoirs of tick-borne pathogens (TBPs). However, limited information is available regarding the prevalence of tick-borne infections in water buffaloes in the Philippines. This study was conducted to identify TBPs harbored by water buffaloes and to characterize these pathogens molecularly. One hundred water buffalo blood samples collected from three areas in Bohol, Visayas region, Philippines were screened for various TBPs using pathogen-specific PCR assays. TBPs were detected in 46% of the samples (39% singly infected, 7% coinfected). The pathogens detected were Anaplasma marginale (29%), Babesia bovis (21%), and B. bigemina (3%). None of the blood samples were positive for Theileria annulata, T. orientalis, and B. ovata. A. marginale infection rates were significantly higher (37.5%) among water buffaloes aged ≤6 years (P = 0.046) than those >6 years old (18.2%) and was detected only in Bulgarian Murrah (36.1%) and US Murrah (25.9%) breeds. Phylogenetic analyses revealed that groEL sequences of A. marginale were 100% identical with isolates from the Philippines (Batangas and Cebu) and China. Two B. bigemina RAP-1a gene sequences were identical to each other and were homologous with previous isolates from Thailand, Indonesia, Uruguay, and the Philippines. Moreover, four B. bovis SBP-2 partial sequences obtained in this study had 92.4-99.7% identities. This study is the first molecular detection and characterization of A. marginale, B. bigemina and B. bovis in water buffaloes in the Visayas region, and the first molecular confirmation of B. bovis infection in water buffaloes in the country. The findings presented in this study may serve as baseline data for crafting effective tick-borne disease surveillance and prevention programs in Bohol and in the Philippines.

First molecular detection of Mycoplasma wenyonii and the ectoparasite biodiversity in dairy water buffalo and cattle in Bohol, Philippines.

Hemoplasmosis caused by Mycoplasma spp. have been associated with major economic losses in the global dairy production. Hemoplasma studies in the Philippines are limited despite its potential impact. This study mainly aimed to detect the presence of hemoplasma species in dairy water buffaloes and cattle and know their ectoparasite biodiversity in Bohol, Philippines. Detection of Mycoplasma spp. was performed using peripheral blood smear examination (PBSE) and standard PCR using whole blood samples collected from 100 dairy water buffaloes and 40 dairy cattle. Available records on the average annual, monthly and daily milk production were compared between PCR-positive and PCR-negative animals. Ectoparasites were manually collected and identified. While PBSE results were all negative, PCR testing showed that 80% (80 water buffaloes and 32 cattle) were positive for Mycoplasma spp. On the other hand, a total of 1436 ectoparasites were collected (609 Haematopinus and 827 Rhipicephalus spp.). DNA sequencing revealed that obtained sequences (193 bp) from 7 animals were 99.5 to 100% similar to registered Mycoplasma wenyonii sequences. The study reports the first molecular characterization of M. wenyonii in the Philippines and probably the first detection in dairy water buffaloes in Southeast Asia.

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.