Infections by Babesia caballi and Theileria equi in Jordanian equids: epidemiology and genetic diversity.

Microscopic diagnosis of equine piroplasmoses, caused by Theileria equi and Babesia caballi, is hindered by low parasitaemia during the latent phase of the infections. However, this constraint can be overcome by the application of PCR followed by sequencing. Out of 288 animals examined, the piroplasmid DNA was detected in 78 (27·1%). Multiplex PCR indicated that T. equi (18·8%) was more prevalent than B. caballi (7·3%), while mixed infections were conspicuously absent. Sequences of 69 PCR amplicons obtained by the 'catch-all' PCR were in concordance with those amplified by the multiplex strategy. Computed minimal adequate model analyses for both equine piroplasmid species separately showed a significant effect of host species and age in the case of T. equi, while in the B. caballi infections only the correlation with host sex was significant. Phylogenetic analyses inferred the occurrence of three genotypes of T. equi and B. caballi. Moreover, a novel genotype C of B. caballi was identified. The dendrogram based on obtained sequences of T. equi revealed possible speciation events. The infections with T. equi and B. caballi are enzootic in all ecozones of Jordan and different genotypes circulate wherever dense horse population exists.

EMERGENCY PREPAREDNESS OF GAMMA SPECTROMETRY LABORATORIES.

Currently, the territorial Radiation Monitoring Network (RMN) of the Czech Republic consists of seven laboratories equipped with gamma spectrometry High Purity Germanium (HPGe) detectors. From 2007 to 2018, five emergency exercises were carried out to test the sample throughput of these facilities and their staff. The main objective was to identify weaknesses and problem areas in the whole process from the moment of obtaining the samples to logging the results into the central RMN database. The long-term aim of these exercises is to optimize emergency response procedures. The most important factor limiting laboratory capacity is the lack of qualified personnel. The exercises showed that in the current state, these laboratories would be able to operate in 12-hour shifts for 14 days and analyze 1700 samples per day. Emergency exercises have highlighted the fact that this type of exercise should be repeated periodically in order to monitor the performance and analytical capabilities of RMN.

Rodents as intermediate hosts of Hepatozoon ayorgbor (Apicomplexa: Adeleina: Hepatozoidae) from the African ball python, Python regius?

Two experimental trials were performed to elucidate the role of rodents in the life cycle of Hepatozoon species using snakes as intermediate hosts. In one trial, two ball pythons, Python regius Shaw, 1802 were force fed livers of laboratory mice previously inoculated with sporocysts of Hepatozoon ayorgbor Sloboda, Kamler, Bulantová, Votýpka et Modrý, 2007. Transmission was successful in these experimentally infected snakes as evidenced by the appearance of intraerythrocytic gamonts, which persisted until the end of trial, 12 months after inoculation. Developmental stages of haemogregarines were not observed in histological sections from mice. In another experimental trial, a presence of haemogregarine DNA in mice inoculated with H. ayorgbor was demonstrated by PCR in the liver, lungs and spleen.

Triploid Colubrid Snake Provides Insight into the Mechanism of Sex Determination in Advanced Snakes.

The advanced snakes (Caenophidia), the important amniote lineage encompassing more than 3,000 living species, possess highly conserved female heterogamety across all families. However, we still lack any knowledge on the gene(s) and the molecular mechanism controlling sex determination. Triploid individuals spontaneously appear in populations of diploid species and can provide an important insight into the evolution of sex determination. Here, we report a case of spontaneous triploidy in a male of the twin-spotted ratsnake (Elaphe bimaculata) with ZZW sex chromosomes. We speculate that as both ZZ and ZZW individuals develop male gonads, the ratio between the number of Z chromosomes and autosomes, and not the presence of the W chromosome in the genome, drives sex determination in the advanced snakes.

A new species of Hepatozoon (Apicomplexa: Adeleorina) from Python regius (Serpentes: Pythonidae) and its experimental transmission by a mosquito vector.

Hepatozoon ayorgbor n. sp. is described from specimens of Python regius imported from Ghana. Gametocytes were found in the peripheral blood of 43 of 55 snakes examined. Localization of gametocytes was mainly inside the erythrocytes; free gametocytes were found in 15 (34.9%) positive specimens. Infections of laboratory-reared Culex quinquefasciatus feeding on infected snakes, as well as experimental infection of juvenile Python regius by ingestion of infected mosquitoes, were performed to complete the life cycle. Similarly, transmission to different snake species (Boa constrictor and Lamprophis fuliginosus) and lizards (Lepidodactylus lugubris) was performed to assess the host specificity. Isolates were compared with Hepatozoon species from sub-Saharan reptiles and described as a new species based on the morphology, phylogenetic analysis, and a complete life cycle.

New species of Choleoeimeria (Apicomplexa: Eimeriidae) from the veiled chameleon, Chamaeleo calyptratus (Sauria: Chamaeleonidae), with taxonomic revision of eimerian coccidia from chameleons.

Coprological examination of 71 samples from a breeding colony of veiled chameleons, Chamaeleo calyptratus Duméril et Duméril, 1851, revealed a presence of two species of coccidia. In 100% of the samples examined, oocysts of Isospora jaracimrmani Modrý et Koudela, 1995 were detected. A new coccidian species, Choleoeimeria hirbayah sp. n., was discovered in 32.4% of samples from the colony. Its oocysts are tetrasporocystic, cylindrical, 28.3 (25-30) x 14.8 (13.5-17.5) microm, with smooth, bilayered, -1 microm thick wall. Sporocysts are dizoic, ovoidal to ellipsoidal, 10.1 (9-11) x 6.9 (6-7.5) microm, sporocyst wall is composed of two plates joined by a meridional suture. Endogenous development is confined to the epithelium of the gall bladder, with infected cells being typically displaced from the epithelium layer towards lumen. A taxonomic revision of tetrasporocystic coccidia in the Chamaeleonidae is provided.

Altitudinal and seasonal differences of tick communities in dogs from pastoralist tribes of Northern Kenya.

Studies regarding the distribution and ecology of ticks in dogs from Eastern Africa are scarce. Our research was based on a long-term screening of ticks parasitising the domestic dogs living with indigenous people around Lake Turkana, Mt. Kulal and Mt. Nyiru areas, Northern Kenya. A total of 9977 ticks were collected from 1464 dogs of all ages and both sexes. Identification was performed using morphological keys and data were analyzed using the Repeated Measures ANOVA, post-hoc Scheffe test and F test, relating independent variables as seasons and regions. Final results were translated to maps using GIS software. Five species of ticks were identified: Rhipicephalus pulchellus, Rhipicephalus sanguineus sensu lato (s.l.), Rhipicephalus armatus, Amblyomma gemma and Hyalomma truncatum. Our results suggest a statistical difference of the tick community structure related to seasonal and altitudinal distribution. Parasitism with R. armatus and R. pulchellus was higher in September-October than in January, whereas, R. sanguineus s.l. was not influenced by the season. Rhipicephalus armatus was present exclusively on dogs living in semi-desert areas, while R. sanguineus s.l. was the dominant species present on the shores of Lake Turkana. Although R. pulchellus was present in the all studied areas, this species had a significantly higher abundance in the afromontane region of Mt. Kulal and montane xeromorphic forest of Mt. Nyiru; these regions are characterized by elevated humidity and cooler climate. Similar geo-climatic distribution is typical also for A. gemma, which was found in dogs exclusively in Mt. Kulal afromontane area. The current work represents the most extensive study performed on the tick community structure of dogs in Eastern Africa. The results showed a relatively limited tick species diversity, with clear seasonal differences and altitudinal distribution.

Quest for the piroplasms in camels: identification of Theileria equi and Babesia caballi in Jordanian dromedaries by PCR.

DNA of two species of piroplasmids was detected in dromedaries during a survey of blood protozoans in Jordan between 2007 and 2009. Ten clinically healthy camels (10%) originating from three Jordanian districts were found, using a PCR assay, to harbor Theileria or Babesia species in their blood and no mix infection was determined. Analysis of the partial 18S rRNA gene sequences of these parasites allowed their unambiguous identification as equine piroplasmids Babesia caballi (n=6) and Theileria equi (n=4). In case of latter species, a novel genotype was found in horses. This first molecular-based species determination of piroplasmids from camels further contributes to the growing evidence of low host specificity of piroplasmids.

Genetic diversity of the class II major histocompatibility DRA locus in European, Asiatic and African domestic donkeys.

The major histocompatibility complex (MHC) genes coding for antigen presenting molecules are the most polymorphic genes in vertebrate genome. The MHC class II DRA gene shows only small variation in many mammalian species, but it exhibits relatively high level of polymorphism in Equidae, especially in donkeys. This extraordinary degree of polymorphism together with signatures of selection in specific amino acids sites makes the donkey DRA gene a suitable model for population diversity studies. The objective of this study was to investigate the DRA gene diversity in three different populations of donkeys under infectious pressure of protozoan parasites, Theileria equi and Babesia caballi. Three populations of domestic donkeys from Italy (N = 68), Jordan (N = 43), and Kenya (N = 78) were studied. A method of the donkey MHC DRA genotyping based on PCR-RFLP and sequencing was designed. In addition to the DRA gene, 12 polymorphic microsatellite loci were genotyped. The presence of Theileria equi and Babesia caballi parasites in peripheral blood was investigated by PCR. Allele and genotype frequencies, observed and expected heterozygosities and F(IS) values were computed as parameters of genetic diversity for all loci genotyped. Genetic distances between the three populations were estimated based on F(ST) values. Statistical associations between parasite infection and genetic polymorphisms were sought. Extensive DRA locus variation characteristic for Equids was found. The results showed differences between populations both in terms of numbers of alleles and their frequencies as well as variation in expected heterozygosity values. Based on comparisons with neutral microsatellite loci, population sub-structure characteristics and association analysis, convincing evidence of pathogen-driven selection at the population level was not provided. It seems that genetic diversity observed in the three populations reflects mostly effects of selective breeding and their different genetic origins.

A survey for piroplasmids in horses and Bactrian camels in North-Eastern Mongolia.

Equine piroplasmosis caused by Babesia caballi and Theileria equi is widespread in Asia. The presence of these haemozoans in Mongolia was previously confirmed in domestic as well as in reintroduced Przewalski horses in which they cause significant pathology. The data on occurrence of piroplasms from Bactrian camels in Asia is lacking. A total of 192 horses, 70 Bactrian camels, and additional 16 shepherd dogs from the Hentiy province were included in our study. No clinical signs typical for piroplasmid infection were observed during the field survey. Microscopic examination revealed the presence of T. equi in blood smears from 67% of examined horses, with camels and dogs being negative. A two step PCR approach was used to detect piroplasms in peripheral blood. In the first "catch all" PCR reaction, amplification of the 496 bp-long fragment of the SSU rRNA gene enabled the detection of Babesia and Theileria spp. Second round multiplex PCR reaction used for species discrimination allowed the amplification of T. equi- and B. caballi-specific 340 bp and 650 bp-long regions of the SSU rRNA, respectively. This assay detected T. equi in 92.7% of horses, while the infections with B. caballi and dual infections were rare. In both PCR setups, camels and dogs were negative indicating that in the studied region, these hosts do not share piroplasms with horses.