A total of 345 faecal samples were collected from domestic, captive and wild birds in rural areas, urban areas and a Zoo in Algeria. Samples were screened for the presence of parasites belonging to the genus Cryptosporidium Tyzzer, 1910 by microscopy and PCR analysis of the small-subunit rRNA (SSU), actin and 60-kDa glycoprotein (gp60) genes. Cryptosporidium spp. were detected in 31 samples. Sequence analysis of SSU and actin genes revealed the presence of C. baileyi Current, Upton et Haynes, 1986 in domestic chicken broilers (n = 12), captive ostriches (n = 4) and a wild mallard (n = 1), and C. meleagridis Slavin, 1955 in a graylag goose (n = 1), chickens (n = 11) and turkeys (n = 2). Twenty-three chicken and two turkey broilers from five farms were positive for cryptosporidia, with an overall prevalence of 2% and 6%, respectively. Both C. meleagridis and C. baileyi were detected in farmed chicken broilers, with a prevalence ranging from 9% to 69%. Farmed turkeys broilers were positive only for C. meleagridis, with a 13% prevalence at the animal level. Subtyping of C. meleagridis isolates at the gp60 locus showed the presence of subtype IIIgA22G3R1 in graylag goose and chicken broilers and IIIgA23G2R1 in chicken and turkey broilers. Infection with cryptosporidia was not associated with any clinical diseases. The results of the present study, which provides the first data on the prevalence of Cryptosporidium spp. in wild birds in Africa, demonstrate the presence of human pathogenic C. meleagridis in both domestic and wild birds in Algeria., Abd Elkarim Laatamna, Nikola Holubová, Bohumil Sak, Martin Kváč., and Obsahuje bibliografii
Pseudorhabdosynochus seabassi sp. n. (Monogenea: Diplectanidae) from the gill filaments of Lates calcarifer Bloch, a marine teleost fish held in floating sea cages in Guangdong Province, China, is described based on morphological observations and molecular data. The shapes of the male copulatory organs (MCO) of Pseudorhabdosynochus spp. were the focus of this study. The typical proximal part of the MCO in most species of Pseudorhabdosynochus is reniform, heavily sclerotized, and divided into four chambers. However, the new species from L. calcarifer has a bulbous proximal region with four concentric layers of apparent muscular origin, instead of a reniform structure with four compartments. This organ is also different in Diplectanum grouperi Bu, Leong, Wong, Woo et Foo, 1999, being sclerotized, cup-shaped, wide proximally with four concentric muscular layers and tubular distally. The 3' terminal portion of the small subunit ribosomal RNA gene (ssrDNA) and the 5' terminal region (domains C1-D2) of the large subunit ribosomal RNA gene (lsrDNA) were used to reconstruct the phylogenetic relationships of P. seabassi and D. grouperi with related taxa utilizing maximum-parsimony and neighbour-joining methods. Phylogenetic analyses unequivocally placed D. grouperi amongst Pseudorhabdosynochus using either ssrDNA or lsrDNA data. All species of Pseudorhabdosynochus (including D. grouperi) used in this study clustered together, inferring monophyly. Based on molecular phylogenetic evidence, we propose that D. grouperi from Epinephelus coioides Hamilton be transferred to Pseudorhabdosynochus as P. grouperi comb. n.
Seven polymorphic microsatellite loci were developed for the planthopper Hyalesthes obsoletus, vector of stolbur 16SrXII-A phytoplasma. The loci have di- or trinucleotide repeat motifs and are highly variable with 10 to 22 alleles per locus. Observed heterozygosity ranged from 0.278 to 0.950 for the 78 individuals genotyped. One locus is sex-linked. No linkage between loci was found. All loci amplified consistently among phylogeographic as well as host-plant related samples and proved highly informative for population genetic studies. and Miriam IMO, Julia LÜNEBURG, Thomas HANKELN, Alfred SEITZ, Jes JOHANNESEN.
Entamoeba moshkovskii Tshalaia, 1941 is prevalent in developing countries and it is considered to be primarily a free-living amoeba, which is morphologically indistinguishable, but biochemically and genetically different from the human infecting, pathogenic Entamoeba histolytica Schaudinn, 1903. The pathogenic potential of this organism is still under discussion. Entamoeba moshkovskii in human stool samples has been reported in different countries such as the United States, Italy, Australia, Iran, Turkey, Bangladesh, India (Pondicherry), Indonesia, Colombia, Malaysia, Tunisia, Tanzania and Brazil, but no data are available about the occurrence of E. moshkovskii in farm animals. This study provides data on the occurrence of E. moshkovskii in pigs in a total of 294 fresh faecal samples collected from five different regions in Kolkata, West Bengal, India. Stool samples were tested by nested PCR using primers targeting SSU rDNA of E. moshkovskii. The amplified PCR products were further confirmed by RFLP technique. Purified nested PCR products were also sequenced and identified via BLAST program run on the NCBI website to confirm species along with their genetic characteristics of the E. moshkovskii isolates. Overall 5.4 % samples were identified as E. moshkovskii positive. Results of this study demonstrate that swine can host E. moshkovskii and should be considered as a potential natural reservoir for E. moshkovskii. However, the occurrence of E. moshkovskii infection in pigs was not statistically associated with their faecal consistency, sex and developmental stage.
In the current study 744 cloacal samples were collected from mallards (Anas platyrhynchos) in the Czech Republic and tested for the presence of influenza virus between 2008 and 2010. Of the total number of 744 mallards tested nine were positive (prevalence 1.2 %) for influenza virus. All the mallards were up to 1.5 years old and the majority (89 %) were killed by hunters.
Species of Hepatozoon Miller, 1908 are vector-borne parasites that infect domestic and wild animals worldwide. Hepatozoon ursi Kubo, Uni, Agatsuma, Nagataki, Panciera et al., 2008 was reported from bears (Ursidae) in Japan and India. The present study represents the first report of infection with H. ursi in Turkish brown bears (Ursus arctos Linnaeus) by microscopic and molecular analysis. Two dead brown bears were found in Uzundere and Pasinler districts of Erzurum. Blood and visceral organ (spleen and liver) samples were delivered to laboratory by the Nature Conservation and National Parks officers. Detected gamonts were evaluated based on morphological features and confirmed as gamonts of H. ursi. The size of gamonts and parasitemia were 8.2 × 3.5 μm (6.9-8.7 × 3.0-3.9 μm; n = 12) and 0.6% (6/1000 leukocytes), respectively. The blood and visceral organ samples were positive for species of Hepatozoon by PCR targeting partial sequence of 18S rDNA. Sequence analysis of newly obtained sequences of H. ursi showed 98.8-100% identity with previously sequenced isolates of H. ursi. Sequences of H. ursi from Erzurum were identical to each other and showed 100% identity with isolates of H. ursi from ticks Ixodes ricinus (Linnaeus), Rhipicephalus turanicus Pomerantzev and Hyalomma marginatum Koch collected from two brown bears in Turkey (GenBank accession numbers MN463021, MN463022, MN905023). Analysis of partial sequences of the 18S rRNA gene of H. ursi showed that Turkish isolates differ in NT substitutions found at three different positions [72 (A→G), 537 (A→G) and 570 (A→T)]. This study provides morphological and molecular data of H. ursi infection in brown bears from two districts of Erzurum, Turkey. Further studies are needed to elucidate whether brown bears have any eco-epidemiologic importance in the life cycle of H. ursi in wildlife.
Parasites of the genus Cryptosporidium Tyzzer, 1910 are one of the most common protistan parasites of vertebrates. Faecal samples from 179 red foxes (Vulpes vulpes [Linnaeus]), 100 grey wolves (Canis lupus Linnaeus), 11 golden jackals (Canis aureus Linnaeus), and 63 brown bears (Ursus arctos Linnaeus) were collected in the Czech Republic, Poland and Slovakia. Samples were examined for the presence of Cryptosporidium spp. using microscopy and PCR/sequence analysis. Phylogenetic analysis based on the small subunit ribosomal RNA (SSU), actin and 60-kDa glycoprotein (gp60) genes using the maximum likelihood method revealed the presence of Cryptosporidium tyzzeri Ren, Zhao, Zhang, Ning, Jian et al., 2012 (n = 1) and C. andersoni Lindsay, Upton, Owens, Morgan, Mead et Blackburn, 2000 (n = 2) in red foxes, C. canis Fayer, Trout, Xiao, Morgan, Lai et Dubey, 2001 (n = 2) and C. ubiquitum Fayer, Santín et Macarisin, 2010 (n = 2) in grey wolves, and C. galli Pavlásek, 1999 in brown bears (n = 1) and red foxes (n = 1). Subtyping of isolates of C. ubiquitum and C. tyzzeri based on sequence analysis of gp60 showed that they belong to the XIId and IXa families, respectively. The presence of specific DNA of C. tyzzeri, C. andersoni and C. galli, which primarily infect the prey of carnivores, is probably the result of their passage through the gastrointestinal tract of the carnivores. Finding C. ubiquitum XIId in wolves may mean broadening the host spectrum of this subtype, but it remains possible this is the result of infected prey passing through the wolf - in this case deer, which is a common host of this parasite. The dog genotype of C. canis was reported for the first time in wolves.
Faecal samples from 162 wild animals were collected from 32 distinct sites of Łęczyńsko-Włodawskie Lakeland (eastern Poland). The presence of Giardia duodenalis (Stiles, 1902) was assessed by a Direct Fluorescence Assay (DFA) and by Polymerase Chain Reaction (PCR) and sequencing of a fragment of the beta-giardin gene. DFA showed the presence of cysts of G. duodenalis in 12 of 162 faecal samples (7%), namely in four wild boars (15%), four foxes (19%), two roe deer (4%), and two wolves (29%). PCR identified 34 of the 162 (21%) samples as positive, including 11 wild boars (41%), five red deer (18%), 11 roe deer (23%), four moose (17%), two wolves (29%) and a single sample from the European badger. Thus, PCR detected a significantly higher number of infection than DFA (P = 0.0005). However, 14 of 34 PCR products could not be sequenced because of their insufficient amount; the low number of cysts, poor conservation of the faeces or presence of PCR inhibitors may have contributed to weak DNA amplification. Sequence analysis of the remaining 20 products showed the presence of assemblage B in wild boars, red deer and roe deer, whereas samples from wolves were identified as assemblage D. This is the first detection of assemblage B in wild boars and deer. As assemblage B has zoonotic potential, wild animals from eastern Poland may act as reservoirs of cysts of G. duodenalis infectious for humans., Krzysztof Stojecki, Jacek Sroka, Simone M. Cacciò, Tomasz Cencek, Jacek Dutkiewicz, Paweł Kusyk., and Obsahuje bibliografii
The community of predators within agroecosystems has the potential to restrict aphid populations, especially early in the season before exponential increases in density and prior to the arrival of specialist natural enemies. Although direct observations of predation, laboratory feeding trials and manipulative field studies have been used to estimate levels of biological control exerted by different species (or potentially negative interactions between them), it is often difficult to extrapolate results to naturally occurring interactions in the field.
Over 100 investigations have utilized gut-content analysis to estimate aphid predation rates by predators. Throughout the last century, gut dissection, which enables the visual identification of aphid body parts, has been used in over 50% of studies although accurate identification and quantification of predation is difficult. Other techniques have included radio-labelling of prey, dissection of faecal samples, electrophoresis, stable isotope analysis and use of polyclonal antisera. In recent studies of invertebrate predation, monoclonal antibodies have been the most frequently applied technique but advances in molecular biology have enabled the detection of species-specific DNA sequences. The use of these applications to quantify predation by aphidophagous predators will be reviewed, with emphasis on potential sources of error and difficulties of quantitative interpretation. Despite the considerable focus currently directed towards molecular approaches, antibody-based techniques are likely to remain an important tool for studying predation rates of pests in the field, especially when antibodies have already been developed. However, the study of multiple predation events within complex generalist predator food webs is only likely through the detection of species-specific DNA sequences using molecular techniques.
In the era of COVID-19 pandemic, organ transplantation programs were facing serious challenges. The lung transplantation donor pool was extremely limited and SARS-CoV-2 viral load assessment has become a crucial part of selecting an optimal organ donor. Since COVID-19 is a respiratory disease, the viral load is thought to be more important in lung transplantations as compared to other solid organ transplantations. We present two challenging cases of potential lung donors with a questionable COVID-19 status. Based on these cases, we suggest that the cycle threshold (Ct) value should always be requested from the laboratory and the decision whether to proceed with transplantation should be made upon complex evaluation of diverse criteria, including the nasopharyngeal swab and bronchoalveolar lavage PCR results, the Ct value, imaging findings and the medical history. However, as the presence of viral RNA does not ensure infectivity, it is still to be clarified which Ct values are associated with the viral viability. Anti-SARS-CoV-2 IgA antibodies may support the diagnosis and moreover, novel methods, such as quantifying SARS-CoV-2 nucleocapsid antigen in serum may provide important answers in organ transplantations and donor selections.