The intestinal protozoan parasite Giardia duodenalis (Lambl, 1859) Kofoid & Christiansen, 1915 [syn. Giardia intestinalis and Giardia lamblia] has emerged as a widespread enteric pathogen in humans and domestic animals. In recent years, G. duodenalis has been found in cattle worldwide and longitudinal studies have reported cumulative prevalence of 100% in some herds. In the present study, we determined the prevalence and genetic characterisation of G. duodenalis in 200 dairy cattle from 10 dairy farms in São Paulo state, Brazil. All faecal specimens were screened for the presence of G. duodenalis using microscopy examination, enzyme immunoassay (EIA) and polymerase chain reaction (PCR). DNA was extracted from faecal samples and G. duodenalis were identified by amplification of the small subunit ribosomal (SSU-rDNA) and glutamate dehydrogenase (GDH) genes followed by restriction fragment length polymorphism (RFLP) or sequencing analysis. Giardia was identified in eight farm locations (80% prevalence). Overall, 15/200 (7.5%) animals were positive for infection, only one of which was a cow. Giardia duodenalis genotype E was present in 14 of the animals tested. Zoonotic genotype AI was present in one positive sample. Genotype E and genotype A represented 93% and 7% of G. duodenalis infections, respectively. This study demonstrates that G. duodenalis infection was prevalent in dairy calves in São Paulo state and that the non-zoonotic genotype E predominates in cattle in this region. Nevertheless, calves naturally infected in Brazil can shed Giardia cysts that can potentially infect humans, and thus, they may represent a public health risk.
Hybrid poplars like Populus ×canadensis or balsam poplar (Populus section Tacamahaca) hybrids have been propagated in Europe since the early 18th century. They replaced many stands of native black poplar on the banks of the major rivers. While spontaneous crosses between hybrid and native black poplars were not considered to be common or of importance in nature, it is shown that such crosses have occurred in Austria, as P. deltoides and section Tacamahaca alleles (PCR-amplified chloroplast and nuclear DNA markers) were found in plants morphologically similar to P. nigra. In the localities studied, a realistic estimate of the introgression rate is between 0 and 10% of plants in a given stand. Female hybrid trees produce viable seed. This shows that hybrid poplars can spread their genes by sexual means, which may result in continuing introgression and consequently, a reduction in genetic diversity and fitness of the endangered P. nigra.
Intestinal microsporidiosis was documented by detecting abundant slightly curved spores (2.9 x 1.2 pm) in the faeces of five of twelve skinks Mabuya perrotetii Duméril et Bibron, 1839 that originated from Ghana. Clinically, the microsporidiosis was characterized by decreased appetite, diarrhea, and weight loss. Histopathological changes consisted of villous atrophy, blunting of mucosa and flattening of individual epithelial cells in the large intestine. The ultrastructure of microsporidian spores was consistent with an Encephalitozoon species. The PCR-RFLP assay and the heleroduplex mobility shift analyses were used to verify that the skink microsporidian is a species of the genus Encephalitozoon Levaditi, Nicolau et Schoen, 1923 and indicate that this microsporidian is not E. hellem, E. intestinalis or a strain of E. cuniculi. The microsporidia in African skink represent an Encephalitozoon species morphologically identical to Encephalitozoon lacerine Canning, 1981.
The aim of the present work was to identify cryptic species in the Anopheles maculipennis and Culex pipiens complexes and to study the genetic structure of the dominant mosquito species Ochlerotatus caspius (Diptera: Culicidae) in the Province of Alessandria close to the vast area untreated rice fields in Piedmont, NW Italy. With the help of PCR-RFLP analysis, four members of the Anopheles maculipennis complex were identified: A. messeae, A. maculipennis, A. sacharovi and A. atroparvus. Only C. pipiens f. molestus was identified in 11 habitats studied in Piedmont. Partial sequences of the cytochrome c oxidase subunit 1 (COI) mitochondrial gene and the second internal transcribed spacer (ITS2) of nuclear ribosomal RNA genes for Italian O. caspius are reported here for the first time. The results indicate that this species diverged from Iranian representatives of this species about one million years ago. The great diversity of mosquito species in Piedmont considerably increases the risk of vector-borne diseases. and Asghar TALBALAGHI, Elena SHAIKEVICH.
Zoonotic cutaneous leishmaniasis (ZCL) is an expanding disease and a public health issue in Iran. In the present study, rate of natural infection of rodent populations with Leishmania was investigated in six endemic foci including 28 villages in Golestan, Esfahan, Yazd, Fars, Khuzestan and Ilam provinces. A total of 593 rodents were captured and identified as Rhombomys opimus (n = 325), Meriones libycus (n = 171), Meriones persicus (n = 27), Tatera indica (n = 37), Nesokia indica (n = 12), Rattus rattus (n = 13) and Mus musculus (n = 8). Microscopic examinations of Giemsa-stained smears showed that 108 out of 593 (18.2%) rodents were infected with Leishmania spp., whereas infection of 186 out of 593 (31.4%) rodents with Leishmania was then confirmed by ITS1-PCR. The highest rate of infection was found in R. opimus (prevalence of 35%) and M. libycus (31%). Based on Restriction Fragment Length Polymorphism (RFLP), 145 (78%) of 186 samples detected as Leishmania DNA were identified as L. major, 8 (4%) samples as L. turanica and 33 (18%) as mixed infection (L. major and L. turanica). Samples from infected rodents were inoculated subcutaneously at tail base of BALB/c mice. In 35 of them, nodules and ulcers containing amastigotes appeared at the inoculation site. The samples prepared from infected rodents were cultured in NNN medium and only two samples were positive. Rhombomys opimus, M. libycus, M. persicus, T. indica and N. indica were confirmed as reservoir hosts of ZCL in the studied regions. Leishmania major infection was usually accompanied L. turanica in naturally infected gerbils (R. opimus and M. libycus) in Golestan, Esfahan and Fars provinces.
Psocids of the genus Liposcelis (Psocoptera: Liposcelididae) are stored product pests that are difficult to identify morphologically. A molecular method based on Restriction Fragment Length Polymorphism (RFLP) of the PCR-amplified 16S rDNA gene was developed for the rapid discrimination of four common species (L. bostrychophila, L. entomophila, L. decolor, and L. paeta). Different developmental stages and populations (P.R. China and Czech Republic) were tested. One DNA fragment of about 500 bp in length was amplified from genomic DNA and the fragment was then digested using the restriction endonuclease DraI. Identification of the relevant banding pattern allowed all the developmental stages and both sexes to be discriminated in the species tested. The banding patterns of L. entomophila from all populations were identical, while the relevant restriction digests and sequence analysis confirmed that the Chinese and Czech populations of L. bostrychophila, L. decolor, and L. paeta differed. In conclusion, PCR-RFLP with one pair of primers (16Sar and 16Sbr) and one restrictive endonuclease, DraI, proved a reliable method for rapidly discriminating the Liposcelis species tested.