Bovine Leukemia Virus molecular detection and associated factors among dairy herd workers in Antioquia, Colombia.

The Bovine Leukemia Virus (BLV) affects mainly cattle, is transmitted by exposure to contaminated biological fluids, and generates lymphomas in 5 % of infected animals. The zoonotic potential of BLV has been studied, and it is currently unknown if it circulates in human workers on dairy herds in Antioquia. Objective: To determine the frequency of BLV detection, the genotypes of the virus, and the factors associated with its detection in workers for dairy herds in Antioquia, Colombia. Through a cross-sectional study in 51 dairy herds, 164 adults were recruited. A peripheral blood sample was collected from each participant for molecular detection of the BLV env and tax genes, and associated factors were explored through bivariate and multivariate mixed Poisson model analyses. The analysis showed that 82 % (134/164) of the participants were men, with an average age of 40. Using qPCR, the constitutive gene GAPDH was amplified to evaluate the presence of amplification inhibitors in the DNA samples. Using nested PCR, the amplification of the env viral gene was obtained in 13 % (22/164) of the total samples analyzed, while all the samples tested negative for tax. The amplicons of the env gene were sequenced, and the identity compatible with BLV was verified by BLAST analysis (NCBI). Using molecular phylogeny analysis, based on maximum likelihood and haplotype network analysis, it was identified that BLV genotype 1 is present in the evaluated population. 16 % (26/164) of the participants reported having ever had an accident with surgical material during work with cattle; this variable was associated with BLV positivity even after adjusting for other variables (PRa =2.70, 95 % CI= 1.01- 7.21). Considering that other studies have reported the circulation of BLV genotype 1 in cattle from this same region and the present report in humans from dairy herds, the results suggest a possible zoonotic transmission of BLV genotype 1 in Antioquia, reinforcing the need to continue investigating to determine the potential role of this virus as an etiological agent of disease in livestock farmers in the department.

Mitochondrial genome comparison and phylogenetic position of Fannia pusio among the Calyptratae flies.

Fannia pusio, the chicken dung fly species, remains unexplored despite its forensic, sanitary, and veterinary importance in the Nearctic and Neotropical regions. In this study, we obtained the complete mitochondrial genome of Fannia pusio for the first time using next-generation sequencing. We compared it with previously published mitogenomes of the genus from the Palearctic region, and its phylogenetic position was studied based on the concatenated protein-coding genes (PCGs) dataset of Calyptratae flies. The circular mitochondrial genome of F. pusio is 16,176 bp in length, with a high A + T content (78.3%), whose gene synteny, codon usage analysis, and amino acid frequency are similar to previously reported Fannia mitogenomes. All PCGs underwent purifying selection except the nad2 gene. Interspecific K2P distances of PCGs of Fannia yielded an average of 12.4% (8.1%-21.1%). The Fannia genus is monophyletic and closely related to Muscidae based on molecular data. Further taxonomic sampling is required to deep into the phylogenetic relationships of the originally proposed species-groups and subgroups within the genus. These results provide a valuable dataset for studying the mitochondrial genome evolution and a resource for the taxonomy and systematics of Fannia.

Molecular surveillance of microbial agents from cattle-attached and questing ticks from livestock agroecosystems of Antioquia, Colombia.

Ticks are obligate ectoparasites and vectors of pathogens affecting health, agriculture, and animal welfare. This study collected ticks from the cattle and questing ticks of 24 Magdalena Medio Antioquia region cattle farms. Genomic DNA was extracted from the specimens (individual or pools) of the 2088 adult ticks collected from cattle and 4667 immature questing ticks collected from pastures. The molecular detection of Babesia, Anaplasma, Coxiella and Rickettsia genera was performed by polymerase chain reaction amplification and subsequent DNA sequencing. In a total of 6755 Rhipicephalus microplus DNA samples, Anaplasma marginale was the most detected with a frequency of 2% (Confidence Interval- CI 1.68-2.36), followed by Babesia bigemina with 0.28% (CI 0.16-0.44), Coxiella spp. with 0.15% (CI 0.07-0.27), and Rickettsia spp. with 0.13% (CI 0.06-0.25). Molecular analysis of the DNA sequences obtained from the tick samples revealed the presence of Coxiella-like endosymbiont and R. felis. These results demonstrated the diversity of microorganisms present in R. microplus ticks predominantly associated with cattle and questing ticks from livestock agroecosystems, suggesting their role as reservoirs and potential biological vectors of these microorganisms on the studied sites. Also, it emphasizes the need to combine acarological surveillance with clinical diagnoses and control strategies on regional and national levels.

Bovine leukemia virus detection in humans: A systematic review and meta-analysis.

To review the available studies on the frequency of detection of the bovine leukemia virus in human samples, a systematic review with meta-analysis of the scientific literature was carried out, including papers published in English, Spanish, and Portuguese in 5 multidisciplinary databases. We collected information from different populations following a detailed and reproducible search protocol in which two researchers verified the inclusion and exclusion criteria. We identified 759 articles, of which only 33 met the inclusion criteria. Analyzed studies reported that the presence of the virus was measured in human samples, such as paraffin-embedded breast tissue and peripheral blood from 10,398 individuals, through serological and molecular techniques. An overall virus frequency of 27% (Ranging between 17 and 37%) was observed, with a high-frequency data heterogeneity between studies. The presence of this virus in different human biological samples suggests the need to investigate further its transmission route to humans and its potential role in developing and progressing diseases.

Hard tick species (Acari: Ixodidae) and infestation in two livestock agroecosystems from Antioquia, Colombia.

Tick infestation affects about 80% of livestock globally while transmitting various pathogens causing high economic losses. This study aimed to determine the degree of tick infestation in two regions, North and Middle Magdalena in Antioquia, Colombia, to identify the ixodid tick species found and the associated risk factors. A cross-sectional study was carried out in 48 farms distributed in six municipalities of Antioquia. Two paddocks and eight bovines per farm were evaluated to estimate tick infestation (adults, nymphs, and larvae). Tick species were identified through a morphological and molecular analysis based on partial sequences of data obtained from DNA molecular markers, two mitochondrial (16S rRNA and COI), and one genomic DNA gene (18S rRNA). A multivariate Poisson regression model was applied to estimate the associated risk factors with ticks in cattle. Rhipicephalus microplus, Amblyomma patinoi and Dermacentor nitens were present in the livestock agroecosystems in the Middle Magdalena region; the highest incidence of tick infestation in cows and paddocks was reported in the municipality of Puerto Triunfo. The livestock agroecosystems in Middle Magdalena were characterized by a higher presence of adult R. microplus in cattle. Larval infestation of R. microplus, followed by D. nitens, was also found in paddocks. The multivariate analysis showed that the origin of cattle was the main risk factor associated with the presence of ticks (i.e., when cattle came from outside the farm). Cattle movement between farms in Middle Magdalena can contribute to the spread of ticks in this region.

Virome analysis of three Ixodidae ticks species from Colombia: A potential strategy for discovering and surveying tick-borne viruses.

Ticks are a group of obligate blood-sucking ectoparasites that play a critical role in transmitting several important zoonotic pathogens that can infect animals and humans. Viruses are part of the tick microbiome and are involved in the transmission of important diseases. Furthermore, the little information on these as etiological agents of zoonoses suggests the need to study these microorganisms. For this reason, in this study, we sought to characterize the virome in Rhipicephalus microplus, Dermacentor nitens, and Rhipicephalus sanguineus s.l., which were collected from different domestic animals in Antioquia, Colombia. RNA sequencing was used for virome characterization in these three tick species, using RNA-dependent polymerase as a marker gene. Forty-eight sequences corresponding to 14 different viruses were identified, some of which were previously identified in the tick's virome. Overall, these data indicate that ticks from domestic animals in cattle farms harbor a wide viral diversity at the local scale. Thus, the metatranscriptomic approach provides important baseline information for monitoring the tick virome and to develop future studies on their biology, host-virus interactions, host range, worldwide distribution, and finally, their potential role as emerging vector-borne agents.

Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia.

Rhipicephalus microplus is recognized as a tick species highly prevalent in cattle, with a wide pantropical distribution that seems to continue spreading geographically. However, its role as a biological vector has been scarcely studied in the livestock context. In this study, a 16S rRNA next-generation sequencing analysis was used to determine bacterial diversity in salivary glands and gut of R. microplus from two contrasting livestock agroecosystems in Antioquia, Colombia. Both the culture-independent approach (CI) and the culture-dependent (CD) approach were complementarily adopted in this study. A total of 341 unique OTUs were assigned, the richness showed to be higher in the Northern than in the Middle Magdalena region, and a high diversity was found at the phylum and genus levels in the samples obtained. With the CI approach, Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria were the most common phylum of bacteria regardless of the organ, or geographic origin of the specimens analyzed. While the relative abundance of bacteria at a phylum level with the CD approach varied between analyzed samples, the data obtained suggest that a high diversity of species of bacteria occurs in R. microplus from both livestock agroecosystems. Bacterial genera such as Anaplasma, Coxiella, and Ehrlichia, recognized for their implications in tick-borne diseases, were also detected, together with endosymbionts such as Lysinibacillus, previously reported as a potential tool for biological control. This information is useful to deepen the knowledge about microbial diversity regarding the relations between endosymbionts and pathogens and could facilitate the future development of epidemiological surveillance in livestock systems.

Metatranscriptomic virome assessment of Rhipicephalus microplus from Colombia.

Ticks (Ixodida) are hematophagous ectoparasites that harbor and transmit diverse species of viruses, some of which cause serious diseases with worldwide veterinary and human health consequences. Rhipicephalus microplus is an important cattle tick in Colombia, where it causes significant economic losses. Despite the importance of this tick, its viral profile is unknown. RNA sequencing was used in this study as a surveillance method for virus detection in R. microplus. Most of the viral origin contigs were assigned to two putative viruses: one chuvirus (Wuhan tick virus 2) and one phlebovirus-like (Lihan tick virus). In addition, viral contigs corresponding to two jingmenviruses previously reported in R. microplus from China and Brazil were detected, as well as a novel putative tymovirus, named here as Antioquia tymovirus-like 1 (ATV-like 1). The presence of some of these viruses across numerous regions in the world could have several explanations, including i) a long-term association between those viruses and R. microplus and ii) a consequence of livestock historical trade. Our results shed new light on the virus diversity of this tick species and provide a basis for further studies on the evolutionary history and pathogenic potential of these interesting viruses.

Morphometric similarity among populations of Rhipicephalus microplus (Acari: Ixodidae) in the north-western region of Colombia.

Rhipicephalus microplus is one of the most widely distributed and economically important ticks able to transmit pathogens to vertebrate hosts which cause major constraints to public and livestock health. A better understanding of their population structure is crucial for the development of more effective control strategies. Modern morphometrics has enabled the quantification of size and shape variations to investigate population differences in anatomical structures. We therefore applied landmark-based and outline-based geometric morphometric approaches to study 85 field-collected adult females of three R. microplus Colombian populations along a distance of 28-67 km using the scutum and the basis capituli. Although size differences were found between some populations, mean shape and metric disparity of the analyzed structures were very similar, resulting in low classification scores (<41%). Thus, both structures support a metapopulation of R. microplus at a microgeographic level. The variance of shape between structures does not co-vary, likely indicating morphological modularity. These data provide first clues to understand the metric variation of R. microplus among natural populations from north-western Colombia.

Phylogenetic analysis of Hepatitis E virus strains isolated from slaughter-age pigs in Colombia.

Hepatitis E virus (HEV) is a zoonotic virus that causes acute hepatitis in humans, and can be transmitted via the fecal-oral route. Pigs are considered to be a reservoir for this infection-mainly where the disease is not endemic. In a previous study conducted in Antioquia, which is a region in Colombia where the production and consumption of pork meat is higher than in the rest of the country, the presence of anti-HEV IgG-type antibodies was reported in slaughter-age pigs. Aiming to identify the HEV genotype circulating in swine, animal liver, and feces samples from five swine cattle slaughterhouses located in six different sub-regions of Antioquia were collected. A nested RT-PCR (nRT-PCR) was used in order to amplify the HEV ORF-1 (170bp) and ORF-2 (348, and 958bp). The amplicons yielded in this study were sequenced, and a molecular phylogeny analysis based on the maximum likelihood method, including HEV sequences reported in several countries, was performed. Phylogeny analysis revealed that HEV amplification fragments from Antioquia's pigs were grouped in three clades within the sub-genotype 3a without a specific geographical structure, and were also genetically related to Japanese and American HEV sequences. This analysis provides the first approach on the genetic diversity and circulation dynamics of HEV in Colombian herds.

[Diagnosis of Strongyloides Stercoralis infection: meta-analysis on evaluation of conventional parasitological methods (1980-2013)]. / Infección por Strongyloides stercoralis: metanálisis sobre evaluación de métodos diagnósticos convencionales (1980-2013).

BACKGROUND: Research on diagnostic methods have strongyloidiasis divergent validity and incomplete by not reporting data on safety, efficiency and performance diagnosis. OBJECTIVE: To assess validity, performance, efficiency and safety of four diagnostic conventional parasitological methods for detection of Strongyloides stercoralis infection in the period 1980-2013. METHODS: Systematic review with meta-analysis, exhaustive and reproducible literature search of six databases. Quality of the articles was assessed and meta-analysis was performed under the random effects model, calculating sensitivity, specificity, likelihood ratios, predictive values, proportion of false results, accuracy, odds ratio and Youden index J and ROC curve using Meta-DiSc(es) and Epidat 3.1. RESULTS: 11 studies with 9,025 individuals were included. Sensitivity of the Baermann method was 72%, positive likelihood ratio (LR+) 228 and negative likelihood ratio (LR-) 0.32. The agar plate culture (APC) had a sensitivity of 89%, LR+ 341 and LR- 0.11. Stool sensitivity was 21%, LR+ 67 and LR- 0.67. Sensitivity of the formol-ether concentration was 48%, LR+ 110 and LR- 0.59. Areas under the ROC curve were 0.999 in Baermann and APC, 0.977 in the stool and 0.829 in formalin-ether concentration; specificity was 100% in all tests. CONCLUSION: The four conventional parasitological methods tested in this study to detect S. stercoralis can be helpful; however, agar plate culture and Baermann method are best suited.

Geometric morphometric analysis of Colombian Anopheles albimanus (Diptera: Culicidae) reveals significant effect of environmental factors on wing traits and presence of a metapopulation.

Anopheles albimanus is a major malaria mosquito vector in Colombia. In the present study, wing variability (size and shape) in An. albimanus populations from Colombian Maracaibo and Chocó bio-geographical eco-regions and the relationship of these phenotypic traits with environmental factors were evaluated. Microsatellite and morphometric data facilitated a comparison of the genetic and phenetic structure of this species. Wing size was influenced by elevation and relative humidity, whereas wing shape was affected by these two variables and also by rainfall, latitude, temperature and eco-region. Significant differences in mean shape between populations and eco-regions were detected, but they were smaller than those at the intra-population level. Correct assignment based on wing shape was low at the population level (<58%) and only slightly higher (>70%) at the eco-regional level, supporting the low population structure inferred from microsatellite data. Wing size was similar among populations with no significant differences between eco-regions. Population relationships in the genetic tree did not agree with those from the morphometric data; however, both datasets consistently reinforced a panmictic population of An. albimanus. Overall, site-specific population differentiation is not strongly supported by wing traits or genotypic data. We hypothesize that the metapopulation structure of An. albimanus throughout these Colombian eco-regions is favoring plasticity in wing traits, a relevant characteristic of species living under variable environmental conditions and colonizing new habitats.

Genetic diversity of Anopheles triannulatus s.l. (Diptera: Culicidae) from northwestern and southeastern Colombia.

Anopheles triannulatus s.l. is a species complex, however in Colombia its taxonomic status is unclear. This study was conducted to understand the level of genetic differentiation or population structure of specimens of An. triannulatus s.l. from northwestern and southeastern Colombia. Cytochrome oxidase subunit I (COI) and internal transcribed spacer (ITS2) sequence analyses suggested high genetic differentiation between the NW and SE populations. A TCS network and Bayesian inference analysis based on 814 bp of COI showed two main groups: group I included samples from the NW and group II samples from the SE. Two main ITS2-polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) patterns were found. Pattern I is present in both the NW and SE, and pattern II is found in the SE specimens. To further elucidate the taxonomic status of An. triannulatus s.l. in Colombia and how these COI lineages are related to the Triannulatus Complex species, the evaluation of immature stages, male genitalia, and additional mitochondrial and nuclear markers will be needed.

Molecular evidence for a single taxon, Anopheles nuneztovari s.l., from two endemic malaria regions in Colombia

To elucidate the Anopheles nuneztovari s.l. taxonomic status at a microgeographic level in four malaria endemic localities from Antioquia and Córdoba, Colombia, fragments of the cytochrome oxidase subunit I (COI) and the white gene were used. The COI analysis showed low genetic differentiation with fixation index (F ST) levels between -0.02-0.137 and Nm values between 3-∞, indicating the presence of high gene flow among An. nuneztovari s.l. populations from the four localities. The COI network showed a single most common haplotype, type 1 (n = 55), present in all localities, as the likely ancestral haplotype. Analysis of the white gene showed that An. nuneztovari s.l. populations from both departments grouped with haplotypes 19 and 20, which are part of lineage 3 reported previously. The results of the present study suggest that An. nuneztovari s.l. is a single taxon in the area of the present study.

Molecular evidence for a single taxon, Anopheles nuneztovari s.l., from two endemic malaria regions in Colombia.

To elucidate the Anopheles nuneztovari s.l. taxonomic status at a microgeographic level in four malaria endemic localities from Antioquia and Córdoba, Colombia, fragments of the cytochrome oxidase subunit I (COI) and the white gene were used. The COI analysis showed low genetic differentiation with fixation index (F(ST)) levels between -0.02-0.137 and Nm values between 3-∞, indicating the presence of high gene flow among An. nuneztovari s.l. populations from the four localities. The COI network showed a single most common haplotype, type 1 (n = 55), present in all localities, as the likely ancestral haplotype. Analysis of the white gene showed that An. nuneztovari s.l. populations from both departments grouped with haplotypes 19 and 20, which are part of lineage 3 reported previously. The results of the present study suggest that An. nuneztovari s.l. is a single taxon in the area of the present study.

A new mtDNA COI gene lineage closely related to Anopheles janconnae of the Albitarsis complex in the Caribbean region of Colombia

An understanding of the taxonomic status and vector distribution of anophelines is crucial in controlling malaria. Previous phylogenetic analyses have supported the description of six species of the Neotropical malaria vector Anopheles (Nyssorhynchus) albitarsis s.l. (Diptera: Culicidae): An. albitarsis, Anopheles deaneorum, Anopheles marajoara, Anopheles oryzalimnetes, Anopheles janconnae and An. albitarsis F. To evaluate the taxonomic status of An. albitarsis s.l. mosquitoes collected in various localities in the Colombian Caribbean region, specimens were analyzed using the complete mitochondrial DNA cytochrome oxidase I (COI) gene, the ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS2) region and partial nuclear DNA white gene sequences. Phylogenetic analyses of the COI gene sequences detected a new lineage closely related to An. janconnae in the Caribbean region of Colombia and determined its position relative to the other members of the complex. However, the ITS2 and white gene sequences lacked sufficient resolution to support a new lineage closely related to An. janconnae or the An. janconnae clade. The possible involvement of this new lineage in malaria transmission in Colombia remains unknown, but its phylogenetic closeness to An. janconnae, which has been implicated in local malaria transmission in Brazil, is intriguing.

Late Pleistocene environmental changes lead to unstable demography and population divergence of Anopheles albimanus in the northern Neotropics.

We investigated the historical demography of Anopheles albimanus using mosquitoes from five countries and three different DNA regions, the mitochondrial cytochrome oxidase subunit I gene (COI), the single copy nuclear white gene and the ribosomal internal transcribed spacer two (ITS2). All the molecular markers supported the taxonomic status of a single species of An. albimanus. Furthermore, agreement between the COI and the white genes suggested a scenario of Pleistocene geographic fragmentation (i.e., population contraction) and subsequent range expansion across southern Central America.

Microgeographic genetic variation of the malaria vector Anopheles darlingi root (Diptera: Culicidae) from Cordoba and Antioquia, Colombia.

Anopheles darlingi is an important vector of Plasmodium spp. in several malaria-endemic regions of Colombia. This study was conducted to test genetic variation of An. darlingi at a microgeographic scale (approximately 100 km) from localities in Córdoba and Antioquia states, in western Colombia, to better understand the potential contribution of population genetics to local malaria control programs. Microsatellite loci: nuclear white and cytochrome oxidase subunit I (COI) gene sequences were analyzed. The northern white gene lineage was exclusively distributed in Córdoba and Antioquia and shared COI haplotypes were highly represented in mosquitoes from both states. COI analyses showed these An. darlingi are genetically closer to Central American populations than southern South American populations. Overall microsatellites and COI analysis showed low to moderate genetic differentiation among populations in northwestern Colombia. Given the existence of high gene flow between An. darlingi populations of Córdoba and Antioquia, integrated vector control strategies could be developed in this region of Colombia.

A new mtDNA COI gene lineage closely related to Anopheles janconnae of the Albitarsis complex in the Caribbean region of Colombia.

An understanding of the taxonomic status and vector distribution of anophelines is crucial in controlling malaria. Previous phylogenetic analyses have supported the description of six species of the Neotropical malaria vector Anopheles (Nyssorhynchus) albitarsis s.l. (Diptera: Culicidae): An. albitarsis, Anopheles deaneorum, Anopheles marajoara, Anopheles oryzalimnetes, Anopheles janconnae and An. albitarsis F. To evaluate the taxonomic status of An. albitarsis s.l. mosquitoes collected in various localities in the Colombian Caribbean region, specimens were analyzed using the complete mitochondrial DNA cytochrome oxidase I (COI) gene, the ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS2) region and partial nuclear DNA white gene sequences. Phylogenetic analyses of the COI gene sequences detected a new lineage closely related to An. janconnae in the Caribbean region of Colombia and determined its position relative to the other members of the complex. However, the ITS2 and white gene sequences lacked sufficient resolution to support a new lineage closely related to An. janconnae or the An. janconnae clade. The possible involvement of this new lineage in malaria transmission in Colombia remains unknown, but its phylogenetic closeness to An. janconnae, which has been implicated in local malaria transmission in Brazil, is intriguing.

Species composition and natural infectivity of anthropophilic Anopheles (Diptera: Culicidae) in the states of Córdoba and Antioquia, Northwestern Colombia

Malaria is a serious health problem in the states of Córdoba and Antioquia, Northwestern Colombia, where 64.4 percent of total Colombian cases were reported in 2007. Because little entomological information is available in this region, the aim of this work was to identify the Anopheles species composition and natural infectivity of mosquitoes distributed in seven localities with highest malaria transmission. A total of 1,768 Anopheles mosquitoes were collected using human landing catches from March 2007-July 2008. Ten species were identified; overall, Anopheles nuneztovari s.l. was the most widespread (62 percent) and showed the highest average human biting rates. There were six other species of the Nyssorhynchus subgenus: Anopheles albimanus (11.6 percent), Anopheles darlingi (9.8 percent), Anopheles braziliensis (6.6 percent), Anopheles triannulatus s.l. (3.5 percent), Anopheles albitarsis s.l. and Anopheles oswaldoi s.l. at < 1 percent; and three of the Anopheles subgenus: Anopheles punctimacula, Anopheles pseudopunctipennis s.l. and Anopheles neomaculipalpusat < 1 percent each. Two species from Córdoba, An. nuneztovari and An. darlingi, were found to be naturally infected by Plasmodium vivax VK247, as determined by ELISA and confirmed by nested PCR. All species were active indoors and outdoors. These results provide basic information for targeted vector control strategies in these localities.