Africanized honeybee population (Apis mellifera L.) in Nicaragua: Forewing length and mitotype lineages.

Various subspecies of Apis mellifera L. were introduced to Central America since colonization 500 years ago. Hybridization increased with the entrance of the Africanized bee in Nicaragua in 1984. Rustic beekeeping activities and numerous feral swarms define the genetic pattern, reflected in phenotypic heterogeneity and remarkable differences in the behaviour of the bee colonies, especially the nest defence. Due to these facts, the question emerge about the degree of Africanization of honeybee colonies in Nicaragua. In this study, we identified Africanized honeybee colonies based on the single character "mean forewing length" and we corroborated our results by determining mitotypes using mtDNA analysis. Morphometric and genetic approaches were realized in three different geographical zones of Nicaragua and related to beehive characteristics and management. Worker bee samples were taken from the inside of 146 hives from 26 apiaries. Abdominal colour as phenotypic character was the first examination, followed by measurement of 1460 right forewings to determine corresponding probability of Africanization. More than 60% of the beehives showed phenotypic heterogeneity and mean forewing length of 8.74 mm (SD 0.16 mm) indicated a high degree of Africanization. Those results provided a selection of 96 worker bees to perform PCR of two worker bees per hive. For mitochondrial DNA analysis 14 samples from sentinel apiaries were added. Three from 61 beehives presented bees with different mtDNA. Throughout, three mitotypes of the African (A) lineage were detected; one mitotype is still unidentified. Mitotype A1 A. mellifera iberiensis was represented by 88 bees and mitotype A4 A. mellifera scutellata by 21 bees. Phylogenetic analysis confirmed the PCR findings. No associations were found between mitotypes, forewing length, beehive characteristics and management. A high degree of Africanization in A. mellifera colonies represented by two predominating mitotypes from the A lineage, prevail in Neotropical Nicaragua, with mitotype A4 predominating at higher altitudes.

Morphotype and haplotype identification of Varroa destructor (Acari: Varroidae), and its importance for apiculture in Nicaragua.

The ectoparasite Varroa (Acari: Varroidae) is considered to be the main pest of honey bees (Apis mellifera L.) in Nicaragua. The aim of this study was to determine morphotypes and mitochondrial haplotypes of the Varroa mites, related to infestation levels in A. mellifera hives in Nicaragua in a cross-sectional study (2013-2016). Samples were taken from 34 sentinel apiaries in five geographical zones; from 713 Varroa females collected during the study, 153 were selected for measurement of various morphometric characters for further classification into morphotypes. The mitochondrial haplotype was assigned to one of the two (Korean or Japanese), using the restriction by SacI of the PCR product of a fragment of the gene CO-I. Morphometric measurement and sequencing revealed the affiliation to the species Varroa destructor with a mean (± SD) body width of 1699.1 ± 60.2 µm and a body length of 1161.1 ± 34.9 µm. Body characters were significantly different among the 34 apiaries. Varroa destructor samples were classified into four morphotypes, with no significant differences in the geographical zones. As GAGCTC SacI enzyme cleavage sequences were not observed, all PCR products were identified as V. destructor Korean haplotype. The CO-I gene nucleotide sequences from two samples V. destructor showed both 100% similarity with the Korean haplotype and 99.8% similarity to the Japanese haplotype. Although the V. destructor mite was identified as a Korean haplotype, host-parasite association in 2 decades has led into a balance without entering into severe losses in the Nicaraguan apiculture.

Leptospira spp in Rodents from Peridomestic Sites in Endemic Regions of Nicaragua.

Leptospirosis is one of the most important zoonoses in tropical countries, including Nicaragua, where it is considered endemic. The aim of this study was to determine the frequency of Leptospira spp in rodents captured from peridomestic sites in leptospirosis endemic regions of Nicaragua. Using live traps, 191 rodents were captured in 2012 and 2013 between April and December. Kidney samples were collected and processed for Leptospira detection from 166 animals by direct culture and isolation. The isolates were tested by PCR for LipL32 and lfb1-F genes specific to pathogenic Leptospira species. The trapping success over all sites was 20.2%, with higher rates of success in rainy season (p < 0.05). Leptospira spp were detected in 22.3% of rodents by direct culture methods. Significant differences (p < 0.01) were found in the frequencies of Leptospira positive rodents per month as well as per region. Of the isolated Leptospira spp, 37.5% were positive for pathogenic species by PCR. The frequency of Leptospira positive rodents by isolation could be used as a predictive indicator for the risk of human leptospirosis in Nicaragua.

Detección de Leptospira spp. en animales y muestras ambientales de áreas peridomésticas en Nicaragua

[RESUMEN]. Objetivo. El objetivo de este estudio fue conocer las características epidemiológicas de la leptospirosis en animales domésticos y en los casos de leptospirosis humana en áreas peridomésticas en Nicaragua entre 2014 y 2016. Métodos. Las muestras se extrajeron en áreas donde se confirmaron casos en humanos utilizando un muestreo no probabilístico en 10 de los 17 departamentos del país. Se incluyeron 112 muestras de orina de animales domésticos, 129 muestras de agua y 69 de tierra para aislar leptospiras en medio Ellinghausen-McCullough-Johnson-Harris (EMJH). Además, se aplicó la prueba de microaglutinación (MAT) en 263 muestras de suero de animales y 88 aislados se analizaron mediante PCR. Resultados. En 32,6% (101/310) de las muestras se aislaron espiroquetas, 23,2% (26/112) se aislaron en la orina de animales domésticos, 47,3% (61/129), en las muestras de agua y 20,3 % (14/69), en las de tierra. El análisis de aislamiento mostró diferencias significativas (P < 0,05) entre los departamentos para los diferentes tipos de muestras, y el aislamiento fue más frecuente en agua que en tierra (OR = 3,49; IC95%: 1,56-7,80). El 14,1% (37/263) de los animales fueron reactores en la prueba de microaglutinación. El serogrupo más frecuente fue Icterohaemorrhagiae (40%). En el análisis con la PCR para identificar leptospiras de las especies patógenas 10,2% (9/88) de los aislamientos fueron positivos. Conclusiones. Esta investigación demuestra que los animales domésticos y el ambiente desempeñan un papel importante en la aparición de brotes de la leptospirosis y confirma el comportamiento endémico de la enfermedad en Nicaragua.

[ABSTRACT]. Objective. The objective of this study was to determine the epidemiological characteristics of leptospirosis in pets and in humans in peridomestic settings in Nicaragua between 2014 and 2016. Methods. The samples were taken in areas where cases were confirmed in humans using non-probabilistic sampling in 10 of the country’s 17 departments. This included 112 urine samples from pets, 129 water samples, and 69 soil samples in order to isolate leptospires in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Furthermore, the microscopic agglutination test (MAT) was applied to 263 samples of animal serum, and 88 isolates were analyzed using PCR. Results. In 32.6% (101/310) of the samples, spirochetes were isolated: 23.2% (26/112) in the pet urine, 47.3% (61/129) in water samples, and 20.3% (14/69) in soil samples. Isolation analysis showed significant differences (p<0.05) between departments for the different types of samples, and isolation was more frequent in water than in soil (OR = 3.49; CI95%: 1.56-7.80). In total, 14.1% (37/263) of the animals were reactors in the microscopic agglutination test. The most frequent serogroup was Icterohaemorrhagiae (40%). PCR analysis to identify pathogenic species of leptospires resulted in 10.2% (9/88) positive isolations. Conclusions. This research demonstrates that pets and environment conditions play an important role in the emergence of outbreaks of leptospirosis, and confirms the endemic behavior of the disease in Nicaragua.

[RESUMO]. Objetivo. Descrever as características epidemiológicas da leptospirose em animais domésticos e em casos de leptospirose humana em áreas peridomiciliares na Nicarágua entre 2014 e 2016. Métodos. As amostras foram coletadas por amostragem não probabilística em áreas com casos confirmados de leptospirose humana em 10 das 17 províncias do país. Foram analisadas 112 amostras de urina de animais domésticos, 129 amostras de água e 69 amostras de solo com o uso do meio de cultura padrão para o isolamento de leptospiras (Ellinghausen-McCullough-Johnson-Harris, EMJH). Além disso, foi realizado o teste de aglutinação microscópica em 263 amostras séricas de animais e 88 isolados foram analisados com a técnica de PCR. Resultados. Em 32,6% (101/310) das amostras foram isoladas espiroquetas, sendo 23,2% (26/112) isoladas na urina de animais domésticos, 47,3% (61/129) nas amostras de água e 20,3% (14/69) nas amostras de solo. Houve diferença significativa (P < 0,05) entre as províncias no isolamento nos diferentes tipos de amostras analisadas, sendo o isolamento mais frequente nas amostras de água que de solo (OR = 3,49; IC95%: 1, 56–7,80). Reatividade no teste de aglutinação microscópica foi observada em 14,1% (37/263) das amostras de animais. O sorogrupo mais frequentemente isolado foi Icterohaemorrhagiae (40%). A técnica de PCR demonstrou que 10,2% (9/88) dos isolados eram positivos para espécies patogênicas de leptospiras. Conclusões. Esta pesquisa demonstra que os animais domésticos e o entorno têm papel importante no surgimento de surtos de leptospirose e confirma o comportamento endêmico da doença na Nicarágua.

[Detection of Leptospire spp. in animals and in environmental samples from peridomestic areas in NicaraguaDetecção de Leptospira spp. em animais e em amostras ambientais de áreas peridomiciliares na Nicarágua]. / Detección de Leptospira spp. en animales y muestras ambientales de áreas peridomésticas en Nicaragua.

OBJECTIVE: The objective of this study was to determine the epidemiological characteristics of leptospirosis in pets and in humans in peridomestic settings in Nicaragua between 2014 and 2016. METHODS: The samples were taken in areas where cases were confirmed in humans using non-probabilistic sampling in 10 of the country's 17 departments. This included 112 urine samples from pets, 129 water samples, and 69 soil samples in order to isolate leptospires in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Furthermore, the microscopic agglutination test (MAT) was applied to 263 samples of animal serum, and 88 isolates were analyzed using PCR. RESULTS: In 32.6% (101/310) of the samples, spirochetes were isolated: 23.2% (26/112) in the pet urine, 47.3% (61/129) in water samples, and 20.3% (14/69) in soil samples. Isolation analysis showed significant differences (p<0.05) between departments for the different types of samples, and isolation was more frequent in water than in soil (OR = 3.49; CI95%: 1.56-7.80). In total, 14.1% (37/263) of the animals were reactors in the microscopic agglutination test. The most frequent serogroup was Icterohaemorrhagiae (40%). PCR analysis to identify pathogenic species of leptospires resulted in 10.2% (9/88) positive isolations. CONCLUSIONS: This research demonstrates that pets and environment conditions play an important role in the emergence of outbreaks of leptospirosis, and confirms the endemic behavior of the disease in Nicaragua.

OBJETIVO: Descrever as características epidemiológicas da leptospirose em animais domésticos e em casos de leptospirose humana em áreas peridomiciliares na Nicarágua entre 2014 e 2016. MÉTODOS: As amostras foram coletadas por amostragem não probabilística em áreas com casos confirmados de leptospirose humana em 10 das 17 províncias do país. Foram analisadas 112 amostras de urina de animais domésticos, 129 amostras de água e 69 amostras de solo com o uso do meio de cultura padrão para o isolamento de leptospiras (Ellinghausen-McCullough-Johnson-Harris, EMJH). Além disso, foi realizado o teste de aglutinação microscópica em 263 amostras séricas de animais e 88 isolados foram analisados com a técnica de PCR. RESULTADOS: Em 32,6% (101/310) das amostras foram isoladas espiroquetas, sendo 23,2% (26/112) isoladas na urina de animais domésticos, 47,3% (61/129) nas amostras de água e 20,3% (14/69) nas amostras de solo. Houve diferença significativa (P < 0,05) entre as províncias no isolamento nos diferentes tipos de amostras analisadas, sendo o isolamento mais frequente nas amostras de água que de solo (OR = 3,49; IC95%: 1,56­7,80). Reatividade no teste de aglutinação microscópica foi observada em 14,1% (37/263) das amostras de animais. O sorogrupo mais frequentemente isolado foi Icterohaemorrhagiae (40%). A técnica de PCR demonstrou que 10,2% (9/88) dos isolados eram positivos para espécies patogênicas de leptospiras. CONCLUSÕES: Esta pesquisa demonstra que os animais domésticos e o entorno têm papel importante no surgimento de surtos de leptospirose e confirma o comportamento endêmico da doença na Nicarágua.

Detección de Leptospira spp. en animales y muestras ambientales de áreas peridomésticas en Nicaragua / Detection of Leptospire spp. in animals and in environmental samples from peridomestic areas in Nicaragua / Detecção de Leptospira spp. em animais e em amostras ambientais de áreas peridomiciliares na Nicarágua

RESUMEN Objetivo El objetivo de este estudio fue conocer las características epidemiológicas de la leptospirosis en animales domésticos y en los casos de leptospirosis humana en áreas peridomésticas en Nicaragua entre 2014 y 2016. Métodos Las muestras se extrajeron en áreas donde se confirmaron casos en humanos utilizando un muestreo no probabilístico en 10 de los 17 departamentos del país. Se incluyeron 112 muestras de orina de animales domésticos, 129 muestras de agua y 69 de tierra para aislar leptospiras en medio Ellinghausen-McCullough-Johnson-Harris (EMJH). Además, se aplicó la prueba de microaglutinación (MAT) en 263 muestras de suero de animales y 88 aislados se analizaron mediante PCR. Resultados En 32,6% (101/310) de las muestras se aislaron espiroquetas, 23,2% (26/112) se aislaron en la orina de animales domésticos, 47,3% (61/129), en las muestras de agua y 20,3 % (14/69), en las de tierra. El análisis de aislamiento mostró diferencias significativas (P < 0,05) entre los departamentos para los diferentes tipos de muestras, y el aislamiento fue más frecuente en agua que en tierra (OR = 3,49; IC95%: 1,56-7,80). El 14,1% (37/263) de los animales fueron reactores en la prueba de microaglutinación. El serogrupo más frecuente fue Icterohaemorrhagiae (40%). En el análisis con la PCR para identificar leptospiras de las especies patógenas 10,2% (9/88) de los aislamientos fueron positivos. Conclusiones Esta investigación demuestra que los animales domésticos y el ambiente desempeñan un papel importante en la aparición de brotes de la leptospirosis y confirma el comportamiento endémico de la enfermedad en Nicaragua.

ABSTRACT Objective The objective of this study was to determine the epidemiological characteristics of leptospirosis in pets and in humans in peridomestic settings in Nicaragua between 2014 and 2016. Methods The samples were taken in areas where cases were confirmed in humans using non-probabilistic sampling in 10 of the country's 17 departments. This included 112 urine samples from pets, 129 water samples, and 69 soil samples in order to isolate leptospires in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Furthermore, the microscopic agglutination test (MAT) was applied to 263 samples of animal serum, and 88 isolates were analyzed using PCR. Results In 32.6% (101/310) of the samples, spirochetes were isolated: 23.2% (26/112) in the pet urine, 47.3% (61/129) in water samples, and 20.3% (14/69) in soil samples. Isolation analysis showed significant differences (p<0.05) between departments for the different types of samples, and isolation was more frequent in water than in soil (OR = 3.49; CI95%: 1.56-7.80). In total, 14.1% (37/263) of the animals were reactors in the microscopic agglutination test. The most frequent serogroup was Icterohaemorrhagiae (40%). PCR analysis to identify pathogenic species of leptospires resulted in 10.2% (9/88) positive isolations. Conclusions This research demonstrates that pets and environment conditions play an important role in the emergence of outbreaks of leptospirosis, and confirms the endemic behavior of the disease in Nicaragua.

RESUMO Objetivo Descrever as características epidemiológicas da leptospirose em animais domésticos e em casos de leptospirose humana em áreas peridomiciliares na Nicarágua entre 2014 e 2016. Métodos As amostras foram coletadas por amostragem não probabilística em áreas com casos confirmados de leptospirose humana em 10 das 17 províncias do país. Foram analisadas 112 amostras de urina de animais domésticos, 129 amostras de água e 69 amostras de solo com o uso do meio de cultura padrão para o isolamento de leptospiras (Ellinghausen-McCullough-Johnson-Harris, EMJH). Além disso, foi realizado o teste de aglutinação microscópica em 263 amostras séricas de animais e 88 isolados foram analisados com a técnica de PCR. Resultados Em 32,6% (101/310) das amostras foram isoladas espiroquetas, sendo 23,2% (26/112) isoladas na urina de animais domésticos, 47,3% (61/129) nas amostras de água e 20,3% (14/69) nas amostras de solo. Houve diferença significativa (P < 0,05) entre as províncias no isolamento nos diferentes tipos de amostras analisadas, sendo o isolamento mais frequente nas amostras de água que de solo (OR = 3,49; IC95%: 1,56-7,80). Reatividade no teste de aglutinação microscópica foi observada em 14,1% (37/263) das amostras de animais. O sorogrupo mais frequentemente isolado foi Icterohaemorrhagiae (40%). A técnica de PCR demonstrou que 10,2% (9/88) dos isolados eram positivos para espécies patogênicas de leptospiras. Conclusões Esta pesquisa demonstra que os animais domésticos e o entorno têm papel importante no surgimento de surtos de leptospirose e confirma o comportamento endêmico da doença na Nicarágua.

Hard ticks (Acari: Ixodidae) of livestock in Nicaragua, with notes about distribution.

We document the species of ticks that parasitize livestock in Nicaragua. The study was based on tick collection on cattle and horses from 437 farms in nine departments. Of 4841 animals examined (4481 cows and 360 horses), 3299 were parasitized, which represent 68 % of the bovines and 67 % of the equines in study: 59 cows and 25 horses were parasitized by more than one species. In addition, 280 specimens of the entomological museum in León were examined. The ticks found on cattle were Rhipicephalus microplus (75.2 % of the ticks collected), Amblyomma mixtum (20.8 %), A. parvum (2.6 %), A. tenellum (0.7 %), A. maculatum (0.7 %). While the ticks collected from the horses were: Dermacentor nitens (41.5 %), A. mixtum (31.7 %), R. microplus (13.8 %), A. parvum (6.5 %), A. tenellum (3.3 %), D. dissimilis (2.4 %) and A. maculatum (0.8 %).

Mesoamerican nephropathy: a neglected tropical disease with an infectious etiology?

An outbreak of unexplained and severe kidney disease, "Mesoamerican Nephropathy," in mostly young, male sugar cane workers emerged in Central America in the late 1990's. As a result, an estimated 20,000 individuals have died, to date. Unfortunately, and with great consequence to human life, the etiology of the outbreak has yet to be identified. The sugarcane fields in Chichigalpa, Chinandega, Nicaragua, have been involved in the outbreak, and during our initial investigation, we interviewed case patients who experienced fever, nausea and vomiting, arthralgia, myalgia, headache, neck and back pain, weakness, and paresthesia at the onset of acute kidney disease. We also observed a heavy infestation of rodents, particularly of Sigmodon species, in the sugarcane fields. We hypothesize that infectious pathogens are being shed through the urine and feces of these rodents, and workers are exposed to these pathogens during the process of cultivating and harvesting sugarcane. In this paper, we will discuss the epidemic in the Chichigalpa area, potential pathogens responsible for Mesoamerican Nephropathy, and steps needed in order to diagnose, treat, and prevent future cases from occurring.