Systematic Review and Meta-analysis of Role of Felids as Intermediate Hosts in the Life Cycle of Neospora caninum Based on Serological Data.

PURPOSE: Neosporosis is an important widespread parasitic infection caused by N. caninum. It infects a wide range of warm-blooded animals as intermediate hosts and dogs as the definitive host. Nevertheless, there are a number of questions regarding the life cycle and epidemiological aspects of N. caninum. Also, the role of felids (domestic and non-domestic) in the life cycle of N. caninum has been little described. Therefore, this study was conducted to evaluate the global prevalence of N. caninum in domestic and wild felids. METHODS: PubMed, Scopus, Google Scholar, Web of Science, and ScienceDirect databases were searched for articles published on the prevalence of N. caninum in felids until Aprill 2, 2022 and the reference lists of retrieved articles were screened. A random-effects meta-analysis model was used to estimate the pooled prevalence and 95% confidence interval. Heterogeneity among studies was evaluated using Cochran's Q and the I2 statistic. RESULTS: After exclusion of irrelevant articles and duplication removal, 30 studies were eligible for quantitative analysis including 20 studies on domestic cats and 10 studies on wild felids. The overall prevalence of neosporosis infection in cats was 15% (95% CI 10-21%) that was significantly higher in wild felids (26%, 95% CI 13-38%) than in domestic cats (11%, 95% CI 6-16%) (P = 0.03). There was no significant difference in pooled prevalence between male and female domestic cats (P = 0.75). Regarding continent, the lowest prevalence of neosporosis infection was in Asia (9%, 95% CI 1-20%) and the highest was in North America (43.6%, 95% CI 33.9-53.2%) and Africa (18%, 95% CI 9-46%). Higher prevalence was obtained when using the NAT with 22% (95% CI 7-37%), compared to the IFAT (17%, 95% CI 9-24%) and ELISA (6%, 95% CI 2-9%) (P = 0.01). CONCLUSION: The findings highlighted the importance of felids as potential intermediate hosts of neosporosis despite the fact that the source of the parasite for these animals was unknown. Further studies should be performed to investigate the role of this top predator (felids) in maintaining both domestic and sylvatic cycles of Neospora caninum.

Insights into the genetic landscape and presence of Cochliomyia hominivorax in the Caribbean.

The New World screwworm, Cochliomyia hominivorax, is a major parasite that causes myiasis in livestock, humans, and other warm-blooded animals in the western hemisphere. There is a permanent biological border that is maintained between Panama and Colombia, as it has been eradicated from North and Central America. However, it still exists in much of the Caribbean and South America causing an estimated annual loss of $3.6 billion dollars in South America alone. Less information is available for C. hominivorax in the Caribbean. Thus, here we examined its presence and genetic landscape in order to gain insights into this fly's distribution in this region. First, through sampling efforts, novel GPS (Global Positioning System) coordinates were collected. Second, the environmental correlates of those presence points were examined. Next, samples were sequenced in order to obtain a pairwise ΦIT genetic distance matrix. And lastly, this matrix was used to create a genetic landscape of divergence. The results of the genetic landscape show flies as more diverse in Trinidad and Tobago and less diverse in the Dominican Republic. This is perhaps due to the proximity of Trinidad to Venezuela and gene flow may be occurring between these two areas. This information will aid in screwworm surveillance and control programs by providing environmental correlates and a view into the distribution of these flies.

Molecular identification of Trypanosoma cruzi in domestic animals in municipalities of the State of Rio Grande do Norte, Brazil.

Trypanosoma cruzi, the etiologic agent of American trypanosomiasis, is a vector-borne zoonotic parasite which has been little studied regarding its infection in domestic animals. In this study, we evaluated the occurrence of natural infection by T. cruzi in farm animals using molecular markers and phylogenetic analysis in blood clot samples of 60 sheep (Ovis aires), 22 goats (Capra hircus), and 14 horses (Equus caballus) in eight municipalities located in an infection risk area in the state of Rio Grande do Norte (RN), Northeast Region of Brazil. Trypanosoma spp. infection was identified by amplifying the rRNA 18S SSU gene in 48.9% of the samples. The SH022 sample showed 99.8% similarity with the Y strain of T. cruzi in phylogeny, grouped in the DTU II clade. Blood clots of sheep, goats, and horses detected T. cruzi kDNA in 28.3% (17/60), 22.7% (5/22), and 15.4% (2/14) of the samples, respectively. These animals were distributed in the three studied mesoregions throughout the state of RN. The identification of natural infection in domestic animals contributes to expand the epidemiological transmission scenario in an area where T. brasiliensis is the main vector.

Leishmania V. braziliensis infection in asymptomatic domestic animals within an endemic region in the Northeast of Brazil.

BACKGROUND: American cutaneous leishmaniasis is a commonly neglected, vector-borne tropical parasitic disease that is a major public health concern in Brazil. Leishmania (Viannia) braziliensis is the main species associated with the disease. Accurate diagnosis is based on epidemiological surveillance, clinical assessment, and laboratory testing. Leishmania (V.) braziliensis has been detected in several wild and synanthropic mammals. Their epidemiological role has not been entirely elucidated. This study aimed to assess potential L. braziliensis infections in asymptomatic domestic animals, by molecular and serological testing in endemic areas, in the metropolitan region of Recife. METHODS: Blood samples and conjunctival fluids were collected from 232 animals (canids, felids, equines, and caprines) for the detection of L. braziliensis using molecular tests (conventional and real-time polymerase chain reaction [PCR and qPCR]). For immunological detection, blood samples from 115 dogs were assessed using enzyme-linked immunosorbent assay. RESULTS: Real-time quantitative PCR showed positive results for blood and conjunctival samples in all investigated species. The results of the blood and conjunctival samples were 68.2% and 26.9% in Canis familiaris, 100% and 41.7% in Felis catus, 77.3% and 30.8% in Equus caballus/Equus asinus, and 50% and 33.3% in Capra hircus samples, respectively. CONCLUSIONS: Results from this study adds valuable information to our understanding of the role of asymptomatic domestic animals, L. braziliensis life cycle, and American cutaneous leishmaniasis in Northeast Brazil.

Morphological abnormalities and multiple mitochondrial clades of Rhipicephalus haemaphysaloides (Ixodida: Ixodidae).

Rhipicephalus haemaphysaloides is endemic in South and Southeast Asia, and it is known to parasitise domestic animals such as cattle, horses, sheep, and dogs. Despite infestation on multiple hosts, little attention has been paid to its morphological and molecular variation. The present study describes local morphological abnormalities for the first time in R. haemaphysaloides from Malaysia, and highlights the presence of three cytochrome c oxidase subunit I (COI) operational taxonomic units with 0.46-9.81% distances within the so-called R. haemaphysaloides in Malaysia, Thailand, and Pakistan.

Geographical distribution of ixodid ticks and tick-borne pathogens of domestic animals in Ethiopia: a systematic review.

BACKGROUND: In Ethiopia, ixodid ticks and associated tick-borne pathogens (TBPs) are of great importance from both a veterinary and public health point of view. This review aimed at compiling available published data on the distribution of ixodid tick species and TBPs in the country. METHODS: A standard review approach was employed using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. Published peer-reviewed articles and theses/dissertations reporting on ixodid ticks and TBPs in Ethiopia were searched using different keywords in many electronic databases including PubMed, Scopus, Web of Science, Google Scholar, African Journals OnLine, and institutional repositories. Articles were screened based on inclusion and exclusion criteria using the PRISMA flowchart. Data were retrieved from eligible articles and recorded in a preformed data record sheet. Descriptive statistics were employed to present data using graphs. QGIS (Quantum GIS) software version 3.4.5 was used to show the distribution of ixodid tick species and TBPs. RESULTS: Overall, 35 articles that met the inclusion criteria were included in this review. Of these, 24 articles report only on ixodid ticks of domestic animals, six articles report only on TBPs in livestock or ticks, and five articles report on both ticks and TBPs in either animals or ticks. Of these studies, 54% were in the Oromia region, while only 3% of studies were in the Benishangul-Gumuz region. The Gambela region lacked studies on ticks and TBPs. At least 19 ixodid tick species have been recorded from different domestic animals including cattle, small ruminants, donkeys, horses, camels, dogs, and cats. Morphological characterization appears to be the sole method of tick species identification in the country. The distribution and abundance of specific tick species depend on geographical locations and agroecological factors. Sixteen molecularly confirmed TBPs have been identified in animal and tick tissue using molecular methods from only four administrative regions, despite the wide distribution of ticks. Among TBPs, five Anaplasma, two Ehrlichia, two Rickettsia, five Theileria, two Babesia, and one Coxiella species are the major pathogens in both livestock and humans. CONCLUSIONS: Many ixodid ticks circulate in a wide geographical zone of Ethiopia. However, the limited reports on TBPs at the country level in general, and the absence of either tick or TBP reports around the border region with neighboring countries in particular, highlights the need for further study.

Chagas Disease Ecology in the United States: Recent Advances in Understanding Trypanosoma cruzi Transmission Among Triatomines, Wildlife, and Domestic Animals and a Quantitative Synthesis of Vector-Host Interactions.

Chagas disease, a neglected tropical disease present in the Americas, is caused by the parasite Trypanosoma cruzi and is transmitted by triatomine kissing bug vectors. Hundreds of vertebrate host species are involved in the ecology of Chagas disease. The sylvatic nature of most triatomines found in the United States accounts for high levels of animal infections but few reports of human infections. This review focuses on triatomine distributions and animal infections in the southern United States. A quantitative synthesis of available US data from triatomine bloodmeal analysis studies shows that dogs, humans, and rodents are key taxa for feeding triatomines. Imperfect and unvalidated diagnostic tools for wildlife complicate the study of animal T. cruzi infections, and integrated vector management approaches are needed to reduce parasite transmission in nature. The diversity of animal species involved in Chagas disease ecology underscores the importance of a One Health approach for disease research and management.

Free-ranging pigs identified as a multi-reservoir of Trypanosoma brucei and Trypanosoma congolense in the Vavoua area, a historical sleeping sickness focus of Côte d'Ivoire.

BACKGROUND: The existence of an animal reservoir of Trypanosoma brucei gambiense (T. b. gambiense), the agent of human African trypanosomiasis (HAT), may compromise the interruption of transmission targeted by World Health Organization. The aim of this study was to investigate the presence of trypanosomes in pigs and people in the Vavoua HAT historical focus where cases were still diagnosed in the early 2010's. METHODS: For the human survey, we used the CATT, mini-anion exchange centrifugation technique and immune trypanolysis tests. For the animal survey, the buffy coat technique was also used as well as the PCR using Trypanosoma species specific, including the T. b. gambiense TgsGP detection using single round and nested PCRs, performed from animal blood samples and from strains isolated from subjects positive for parasitological investigations. RESULTS: No HAT cases were detected among 345 people tested. A total of 167 pigs were investigated. Free-ranging pigs appeared significantly more infected than pigs in pen. Over 70% of free-ranging pigs were positive for CATT and parasitological investigations and 27-43% were positive to trypanolysis depending on the antigen used. T. brucei was the most prevalent species (57%) followed by T. congolense (24%). Blood sample extracted DNA of T. brucei positive subjects were negative to single round TgsGP PCR. However, 1/22 and 6/22 isolated strains were positive with single round and nested TgsGP PCRs, respectively. DISCUSSION: Free-ranging pigs were identified as a multi-reservoir of T. brucei and/or T. congolense with mixed infections of different strains. This trypanosome diversity hinders the easy and direct detection of T. b. gambiense. We highlight the lack of tools to prove or exclude with certainty the presence of T. b. gambiense. This study once more highlights the need of technical improvements to explore the role of animals in the epidemiology of HAT.

Assessment of the role played by domestic animals in jigger infection in Kandara sub-county, Kenya (case control study).

INTRODUCTION: tungiasis is an ectoparasitosis caused by penetration of female sand flea, Tunga penetrans, into the skin of the susceptible animal and the consequent hypertrophy of the parasite. The objective of this study was to assess the association between domestic animals and jigger infection among the residents of Kandara sub-county in central Kenya. METHODS: this was a case-control study that involved 776 individuals. Half of this number entailed case group who were jigger infected while the other half was the control, composed of jigger free participants. Structured questionnaires were, administered among the heads of the households to which the participants belonged to gather information concerning the animals they kept. Univariate analysis was, applied. RESULTS: in this study, there were significant differences in age (P=0.008) between the two groups. Disparities in source of income (P<0.001) and level of education (P<0.001) came out as very significant factors in jigger infection. The case group was 10 times more likely to keep dogs than the control(9.6; 95% CI, 5.9-15.6). Case group was also 7 times more likely to rear chicken in comparison to the control (6.6; 95%, 4.2-10.4). The case group was 12 times more likely to let dogs loose in the compound in comparison to the control (12.1: 95%, 5.9-24.5). When compared to the control, this group was also 17 times more likely to keep chicken inside their houses (16.7: 95% CI, 6.8-35.9). Conclusion: there is a very high association between domestic animals and occurrence of tungiasis in Kandara sub-county.

A cross-sectional survey of hard ticks and molecular characterization of Rhipicephalus microplus parasitizing domestic animals of Khyber Pakhtunkhwa, Pakistan.

BACKGROUND: In tropical and subtropical countries, tick infestation causes major public health problems and considerable financial losses to the livestock industry. This study was aimed to assess the species composition of richness and analyze the phylogeny of Rhipicephalus microplus in the District Bannu of Khyber Pakhtunkhwa, Pakistan. METHODS: Collected ticks were identified morphologically and DNA extracted from R. microplus was amplified and subjected to sequencing. RESULTS: A total of 3,600 animals were examined among them 1,494 animals were found to be infested with ticks, including 669 cows, 476 buffaloes, 163 goats, and 186 sheep (p = 0.001). Tick infestation was significantly high (43.58%) in animals of age group (<1 year) (p-value = 0.027). Female animals were more (44.05%) infested with ticks than males (34.43%) (p = 0.001). The intensity of infestation was significantly higher in summer (77.49%) (p = 0.001). A total of 5,557 ticks were collected comprising three genera and six species. R. microplus was predominantly prevalent (n = 1,474; 26.52%), followed by Rhipicephalus annulatus (n = 1,215; 21.86%), Hyalomma anatolicum (n = 1,139; 20.49%), Hyalomma marginatum (n = 1,086; 19.54%), and Rhipicephalus turanicus (n = 761; 13.69%), while the least common was Haemaphysalis aciculifer (n = 80; 1.43%) (p = 0.001). Morphologically identified R. microplus species were also analyzed genetically by using two genetic markers 16S ribosomal RNA (16S rRNA) and internal transcribed spacer 2 (ITS2) genes. The phylogenetic study revealed that R. microplus is genetically diversified and clustered in clade B with R. microplus species from China, India, and Pakistan. CONCLUSION: Ticks infestation was significantly correlated with various factors including age, sex, season, and animal type. R. microplus genetically resembled species reported from India and China. However, major knowledge gaps concerning various species of ticks exist and many areas are still unexplored in Pakistan. Therefore, it is necessary to explore the epidemiological and molecular aspects of various tick species in other regions of southern Khyber Pakhtunkhwa.

Herd-level contamination of Neospora caninum, Toxoplasma gondii and Brucella in milk of Iranian dairy farms.

The bulk milk examination is a reliable screening tool for monitoring the quality of milk in the farms. The infection to Neospora caninum, Toxoplasma gondii and Brucella sp. Was evaluated in bulk milk samples of dairy farms in Hamedan province, West part of Iran. All the dairy farms (n = 149) were examined for N. caninum, T. gondii and Brucella infections using milk ring test (MRT), microbiology, serology (Enzyme-linked Immunosorbent Assay), and molecular techniques. Based on molecular methods, Brucella-infection was negative in all farms; while, 55 %, 5.4 % and 2.7 % of samples were positive for N. caninum, T. gondii and mix infection, respectively. The highest Neospora-infection was detected in the farms with history of abortion in fall and winter. There was significant association between Neospora-infection and the presence of dogs and rodents in the farms, herd size, and age of the animals. Also, a significant association was seen between Toxoplasma-infection and the presence of cats and rodents in the farms, as well as age of the animals. Average total bacterial count (TBC) was calculated 1.14 × 106±1.1 × 106. The highest TBC was in the farms from Central locations of studied area (5.7 × 106±2.24 × 106), farms with more than 120 animals (7.9 × 106±2.8 × 106), and farms with ≥50-months age (1.74 × 106±6.3 × 105) in spring and summer (6.9 × 106±3.7 × 106). The number of somatic cells was estimated between 1 × 104 and 2 × 106 (Average = 4.2 × 105±3.39 × 105). The current study was a comprehensive evaluation of Neospora, Toxoplasma and Brucella infections in milk samples of Iranian dairy farms for the first time. Neospora-infection is responsible for economic losses in the region. Health education and milk pasteurization are so helpful for inhibiting the milk borne diseases. To reduce the risk factors, predict and design the appropriate schemes like redundant of heterogeneous animals are recommended.

Prevalence and associated risk factors of Leishmania infection among immunocompetent hosts, a community-based study in Chiang Rai, Thailand.

BACKGROUND: Leishmaniasis is an emerging infectious disease reported in the north and south of Thailand of which patients with HIV/AIDS are a high risk group for acquiring the infection. A lack of information regarding prevalence, and the risk association of Leishmania infection among asymptomatic immunocompetent hosts needs further investigation. Information on potential vectors and animal reservoirs in the affected areas is also important to control disease transmission. METHODS: An outbreak investigation and a cross-sectional study were conducted following one index case of cutaneous leishmaniasis (CL) caused by L. martiniquensis in an immunocompetent male patient reported in August 2015, Chiang Rai Province, Thailand. From September to November 2015, a total of 392 participants at two study areas who were related to the index case, 130 students at a semi-boarding vocational school and 262 hill tribe villagers in the patient's hometown, were recruited in this study. The nested internal transcribed spacer 1-PCR (ITS1-PCR) was performed to detect Leishmania DNA in buffy coat, and nucleotide sequencing was used to identify species. Antibody screening in plasma was performed using the Direct Agglutination Test (DAT), and associated risk factors were analyzed using a standardized questionnaire. Captured sandflies within the study areas were identified and detected for Leishmania DNA using nested ITS1-PCR. Moreover, the animal reservoirs in the study areas were also explored for Leishmania infection. RESULTS: Of 392 participants, 28 (7.1%) were positive for Leishmania infection of which 1 (4.8%) was L. martiniquensis, 12 (57.1%) were L. orientalis and 8 (38.1%) were Leishmania spp. Of 28, 15 (53.6%) were DAT positive. None showed any symptoms of CL or visceral leishmaniasis. Risk factors were associated with being female (adjusted odds ratio, AOR 2.52, 95%CI 1.01-6.26), increasing age (AOR 1.05, 95%CI 1.02-1.08), having an animal enclosure in a housing area (AOR 3.04, 95%CI 1.13-8.22), being exposed to termite mounds (AOR 3.74, 95%CI 1.11-12.58) and having domestic animals in a housing area (AOR 7.11, 95%CI 2.08-24.37). At the semi-boarding vocational school, six Sergentomyia gemmea samples were PCR positive for DNA of L. orientalis and one S. gemmea was PCR positive for DNA of L. donovani/L. infantum. Additionally, one Phlebotomus stantoni was PCR positive for DNA of L. martiniquensis, and one black rat (Rattus rattus) was PCR positive for DNA of L. martiniquensis. CONCLUSION: This information could be useful for monitoring Leishmania infection among immunocompetent hosts in affected areas and also setting up strategies for prevention and control. A follow-up study of asymptomatic individuals with seropositive results as well as those with positive PCR results is recommended.

Zoonotic enteric parasites in Mongolian people, animals, and the environment: Using One Health to address shared pathogens.

BACKGROUND: Cryptosporidium spp. and Giardia duodenalis are important zoonotic enteric pathogens of One Health concern for humans, animals, and the environment. For this study, we investigated parasite prevalence and risk factors among rural, peri-urban, and urban households and environments of Mongolia. METHODS: This cross-sectional study implemented a household risk factor survey at 250 home sites along with sample collection from humans, animals, flies, and drinking water. Multiplex real-time PCR analysis was conducted to look for Cryptosporidium spp. and/or Giardia duodenalis within household samples. RESULTS: Lab analysis found one or both zoonotic parasites at 20% of the participating households (51/250). Human samples had a parasite prevalence of 6.4% (27/419), domestic animals at 3.3% (19/570), pooled filth flies at 14.8% (17/115), and drinking water samples at 2% (5/250). Parasite presence at the household was significantly associated with a household's use of an improved drinking water source (OR 0.27; CI 0.12-0.61; p = < 0.01), having an indoor handwashing site (OR 0.41; CI 0.19-0.92; p = 0.03), domestic animal ownership (OR 2.40; CI 1.02-5.65; p = 0.05), and rural location (OR 0.50; CI 0.25-0.98; p = 0.04). Household use of an improved drinking water source remained significant in the multivariate model (OR 0.16; CI 0.04-0.68; p = 0.01). CONCLUSION: In Mongolia, public and veterinary health are intertwined, particularly for rural herding households. Increased access to safe water, sanitation and hygiene infrastructure could help prevent further transmission of zoonotic enteric parasites. Public health interventions, policy and messaging should utilize a One Health framework employing joint leadership from local human and animal health sectors.

A review of the occurrence and clinical consequences of protozoan infections in carnivorous fur farm animals.

INTRODUCTION AND OBJECTIVE: The aim was to review available data concerning the occurrence of protozoan parasites affecting carnivorous fur farm animals, such as: American mink, blue and silver foxes, and raccoon dogs. Although, many protozoan pathogens have been isolated in wild fur animals, some are still not recognised as relevant to particular fur farm animal species. Protozoans that have been isolated as causative agents of clinical cases and reported in these animals are: Cryptosporidium spp., Eimeria spp., Isospora spp., Leishmania infantum, Neospora caninum, Sarcocystis spp., and Toxoplasma gondii. Unfortunately, in most reports, neither clinical symptoms nor detailed pathogenicity and clinical pathology are satisfactory discriminated. This data is essential for preparing treatment and preventive measure protocols. Additionally, attempts to better understand the infections might be useful in the case of outbreaks. RESULTS: The results of the literature research revealed that some detected infections may have zoonotic potential, and are quite likely to be transmitted in both directions. The role of vectors, e.g. small rodents, birds, insects (sand-flies), or pets (cats and dogs), which might be involved in the transmission of various parasitic infections, is also discussed. The summarized list of protozoans involved can be used for further studies on the health and welfare aspects of fur farm animal breeding and public health issues.

Molecular identification and subtype distribution of Blastocystis sp. in farm and pet animals in Turkey.

A total of 1340 fresh fecal samples from farm and pet animals in Central Anatolia and the Middle Black Sea Region of Turkey were investigated using a PCR assay targeting the SSU rRNA of Blastocystis sp. An overall Blastocystis sp. prevalence of 19.4% (183/940) was found in farm animals, including cattle, sheep, water buffaloes, and chickens. Fecal samples of dogs, cats, and horses were negative. The highest prevalence of Blastocystis sp. was found in sheep (38.2%) among the farm animals. The SSU rRNA sequence analysis revealed two animal-specific subtypes, including ST10 in cattle and sheep and ST14 in water buffaloes. The zoonotic subtype ST7 was identified in chickens. Our results indicated a high prevalence of animal-specific subtypes in livestock and zoonotic subtype ST7 in chickens, highlighting the potential risk of chickens for zoonotic transmission of Blastocystis in the research area. This study is the first large-scale evaluation of Blastocystis in animal hosts in Turkey, and contributes to the molecular epidemiology and genetics of Blastocystis. Our results should be considered by authorities as an indication of the zoonotic importance of Blastocystis sp. and the need for surveillance in public health intervention programs.

Interactive effects of biological, human and environmental factors on tick loads in Boran cattle in tropical drylands.

BACKGROUND: Tick-borne diseases (TBDs) are a serious threat to humans, wildlife and livestock, and cause severe economic losses in many tropical drylands. The effective control of TBDs has been constrained by limited understanding of what determines tick loads in animals. We tested interactive effects of several biological factors (sex, age and body condition), one environmental factor (rainfall) and one human factor (management type) on tick loads in animals. METHODS: We collected ticks on animals at four sampling sites in the semi-arid savanna area of Laikipia County, Kenya, of which two are commercial ranches and the other two are open pastoral grazing areas. From 2017 to 2019, we collected a total of 2038 ticks from 619 domestic animals from various cattle and camel herds and from 79 tranquilised wild animals. RESULTS: Generally, wild herbivores (zebras, rhinos and elephants) had higher tick loads than domestic animals. As 83% of the tick samples were taken from Boran cattle, we analysed tick load in these cattle in more detail. Boran cattle had high tick loads in the wet season, especially those animals in poor condition. No differences were found between female and male cattle, regardless of season. The calves had high tick loads during both the wet and dry seasons, whereas the sub-adult and adult cattle had less ticks during the dry season. Cattle on the intensively managed ranches had lower tick load than those in the transhumant management system. CONCLUSION: These findings highlight the importance of establishing effective control of ticks on domestic animals in transhumant management systems as tick loads were high on these animals in both the wet and dry season.

Development of a loop-mediated isothermal amplification assay based on RoTat1.2 gene for detection of Trypanosoma evansi in domesticated animals.

The early containment of trypanosomosis depends on early, sensitive, and accurate diagnosis in endemic areas with low-intensity infections. The study was planned to develop a simple read out loop-mediated isothermal amplification (LAMP) assay targeting a partial RoTat1.2 VSG gene of Trypanosoma evansi with naked eye visualization of LAMP products by adding SYBR® Green I dye. The visual results were further confirmed with those of agarose gel electrophoresis, restriction enzyme digestion of LAMP products with AluI, and sequencing of the PCR products using LAMP outer primers. The LAMP primers did not show cross reactivity and non-specific reactions with regional common hemoparasitic DNA revealing high specificity of the assay. The threshold sensitivity level of the LAMP assay was determined to be 0.003 fg compared to 0.03 fg RoTat1.2 amplified DNA fragments of T. evansi by PCR assay. Moreover, assessment of 500 blood samples collected from unhealthy domestic animals in field suspected for various hemoparasitic infections was carried out for the presence of T. evansi by microscopy, RoTat1.2 VSG PCR, and LAMP assay. LAMP could detect T. evansi in 36 samples, while PCR and microscopy could detect 33 and 12 samples, respectively. All the samples positive by microscopy and PCR were also confirmed positive by the LAMP assay. The current LAMP assay has appealing point of care characteristics to visually monitor the results, lessen the need of post DNA amplification procedure, and enable this method to be applied as a rapid and sensitive molecular diagnostic tool in under resourced laboratories and field setup.

Correlates of Variation in Guinea Worm Burden among Infected Domestic Dogs.

The Guinea Worm Eradication Program has been extraordinarily successful-in 2019, there were 53 human cases reported, down from the estimated 3.5 million in 1986. Yet the occurrence of Guinea worm in dogs is a challenge to eradication efforts, and underlying questions about transmission dynamics remain. We used routine surveillance data to run negative binomial regressions predicting worm burden among infected dogs in Chad. Of 3,371 infected dogs reported during 2015-2018, 38.5% had multiple worms. A multivariable model showed that the number of dogs in the household was negatively associated with worm burden (adjusted incidence rate ratio [AIRR] = 0.95, 95% CI: 0.93-0.97, P < 0.0001) after adjusting for dog age (AIRR = 0.99, 95% CI: 0.96-1.01, P > 0.1). This could relate to the amount of infective inocula (e.g., contaminated food or water) shared by multiple dogs in a household. Other significant univariable associations with worm burden included dog history of Guinea worm infection (IRR = 1.30, 95% CI: 1.18-1.45) and dog owners who were hunters (IRR = 0.78, 95% CI: 0.62-0.99, P < 0.05) or farmers (IRR = 0.83, 95% CI: 0.77-0.90, P < 0.0001). Further analysis showed that the number of dogs in the household was significantly and positively correlated with nearly all other independent variables (e.g., owner occupation: farmer, fisherman, or hunter; dog age, sex, and history of Guinea worm). The associations we identified between worm burden and dogs per household, and dogs per household and owner characteristics should be further investigated with more targeted studies.

A novel fluorescence immunochromatographic assay strip for the diagnosis of schistosomiasis japonica.

BACKGROUND: Schistosomiasis japonica is a severe zoonosis. Domestic animals are the primary source of infection and play an important role in disease transmission. Surveillance and diagnosis play key roles in schistosomiasis control; however, current techniques for the surveillance and diagnosis of the disease have limitations. In this study, we developed a novel fluorescence immunochromatographic assay (FICA) strip to detect anti-Schistosoma japonicum antibodies in host serum. METHODS: A FICA strip was developed for the diagnosis of Schistosoma japonicum in domestic animals. Streptococcus protein G (SPG) and soluble egg antigen (SEA) were transferred onto a nitrocellulose (NC) membrane to form the control line (C) and the test line (T), respectively. With fluorescence activity as well as binding activity to multispecies IgG, the recombinant protein rSPG-RFP was expressed and employed as an antibody indicator in the FICA strips. RESULTS: The dual gene fusion plasmid was verified by PCR and restriction enzyme digestion. The expressed recombinant protein was 39.72 kDa in size, which was consistent with the predicted molecular weight. The western blot results showed binding activity between rSPG-RFP and IgGs from different hosts. Fluorescence microscopy also showed the fluorescence activity of the protein present. The affinity constant (Ka) values of rSPG-RFP with rabbit, donkey, mouse and goat IgG were 1.9 × 105, 4.1 × 105, 1.7 × 105 and 5.4 × 105, respectively. Moreover, based on the recombinant protein, the test strip for detecting S. japonicum in buffaloes could distinguish positive from negative serum. The lower limit of detection of the FICA strip was 1:10,000. Compared with ELISA, the FICA strips exhibited similar results in the diagnosis of infection in clinical bovine serum samples, with a kappa value of 0.9660 and P < 0.01. The cross-reactivities of the FICA strips with Haemonchus contortus and Schistosoma turkestanicum (30.15% and 91.66%, respectively) were higher than those of ELISA (26.98% and 87.5%, respectively). CONCLUSIONS: Based on the rSPG-RFP protein that we developed, strip detection can be completed within 15 min. Heightened sensitivity allows the strip to accurately identify schistosome antibodies in serum. In conclusion, this method is convenient, feasible, rapid and effective for detecting S. japonicum.