Trichinella spp., are amongst the most widespread parasitic nematodes, primarily live in the muscles of a wide range of vertebrate animals and humans. Human infection occurs by ingestion of raw or undercooked meat containing Trichinella larvae. Accurate diagnosis of Trichinella spp. infection in domestic animals is crucial for the effective prevention and control of human trichinellosis. In the present study, a simple, rapid and accurate diagnostic assay was developed combining recombinase polymerase amplification and a lateral flow strip (LF-RPA) to detect Trichinella spp. infection. The LF-RPA assay targets Trichinella spp. mitochondrial small-subunit ribosomal RNA (rrnS) gene and can detect as low as 100 fg DNA of Trichinella strains, which was approximately 10 times more sensitive than a conventional PCR assay. The LF-RPA assay can be performed within 10–25 min, at a wide range of temperatures (25–45°C) and showed no cross-reactivity with DNA of other parasites and related host species of Trichinella. The performance of the LF-RPA assay in the presence of high concentration of PCR inhibitor was better than that of a conventional PCR assay. Results obtained by LF-RPA assay for the detection of experimentally infected mice were comparable to the results obtained by using a conventional PCR, achieving 100% specificity and high sensitivity. These results present the developed LF-RPA assay as a new simple, specific, sensitive, rapid and convenient method for the detection of Trichinella infection in domestic animals.

Background: Mitochondrial genomes provide a rich source of molecular variation of proven and widespread utility in molecular ecology, population genetics and evolutionary biology. The tapeworm genus Taenia includes a diversity of tapeworm parasites of significant human and veterinary importance. Here we add complete sequences of the mt genomes of T. multiceps, T. hydatigena and T. pisiformis, to a data set of 4 published mtDNAs in the same genus. Seven complete mt genomes of Taenia species are used to compare and contrast variation within and between genomes in the genus, to estimate a phylogeny for the genus, and to develop novel molecular markers as part of an extended mitochondrial toolkit. Results: The complete circular mtDNAs of T. multiceps, T. hydatigena and T. pisiformis were 13,693, 13,492 and 13,387 bp in size respectively, comprising the usual complement of flatworm genes. Start and stop codons of protein coding genes included those found commonly amongst other platyhelminth mt genomes, but the much rarer initiation codon GTT was inferred for the gene atp 6 in T. pisiformis. Phylogenetic analysis of mtDNAs offered novel estimates of the interrelationships of Taenia. Sliding window analyses showed nad 6, nad 5, atp 6, nad 3 and nad 2 are amongst the most variable of genes per unit length, with the highest peaks in nucleotide diversity found in nad 5. New primer pairs capable of amplifying fragments of variable DNA in nad 1, rrn S and nad 5 genes were designed in silico and tested as possible alternatives to existing mitochondrial markers for Taenia. Conclusions: With the availability of complete mtDNAs of 7 Taenia species, we have shown that analysis of amino acids provides a robust estimate of phylogeny for the genus that differs markedly from morphological estimates or those using partial genes; with implications for understanding the evolutionary radiation of important Taenia. Full alignment of the nucleotides of Taenia mtDNAs and sliding window analysis suggests numerous alternative gene regions are likely to capture greater nucleotide variation than those currently pursued as molecular markers. New PCR primers developed from a comparative mitogenomic analysis of Taenia species, extend the use of mitochondrial markers for molecular ecology, population genetics and diagnostics.

Autoimmune diseases (ADs) are featured by body's immune responses being directed towards its own specific target organs or multiple organ systems, causing persistent inflammation and consequent tissue damage. miRNAs are small noncoding RNAs in a size of approximately 22 nt that play important regulatory roles in many organisms by cleavage or translational inhibition of targeted mRNAs. Many miRNAs are reported to be differentially expressed in ADs and may play a pivotal role in regulating immune responses and autoimmunity. In this review, current research progress in the miRNAs in ADs was elucidated.

Background: In recent years, surveys of Toxoplasma gondii infection in dogs have been reported worldwide, including China. However, little is known about the prevalence of T. gondii in pet dogs in Northwest China. In the present study, the prevalence of T. gondii in pet dogs in Lanzhou, China was investigated using the modified agglutination test (MAT). Results: In this survey, antibodies to T. gondii were found in 28 of 259 (10.81%) pet dogs, with MAT titers of 1:20 in 14 dogs, 1:40 in nine, 1:80 in four, and 1:160 or higher in one dog. The prevalence ranged from 6.67% to 16.67% among dogs of different ages, with low rates in young pet dogs, and high rates in older pet dogs. The seroprevalence in dogs >3 years old was higher than that in dogs ≤1 years old, but the difference was not statistically significant (P > 0.05). The seroprevalence in male dogs was 12.50% (17 of 136), and in female dogs it was 8.94% (11 of 123), but the difference was not statistically significant (P > 0.05). Conclusions: A high prevalence of T. gondii infection was found in pet dogs in Lanzhou, Northwest China, which has implications for public health in this region. In order to reduce the risk of exposure to T. gondii, further measures and essential control strategies should be carried out rationally in this region.

Background: Cystic echinococcosis (CE) is a serious tapeworm infection caused by Echinococcus granulosus (sensu lato) which infects a wide range of animals and humans worldwide. Despite the millions of livestock heads reared in Pakistan, only a few reports on CE prevalence and even fewer on the genetic diversity are available for the country. Meanwhile, the available reports on the genetic diversity are predominantly based on short sequences of the cox1 gene. Methods: To close this knowledge gap, this study was designed to investigate the genetic diversity and population structure of Echinococcus spp. in Pakistan using the complete mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes. Results: Based on BLAST searches of the generated cox1 and nad1 gene sequences from a total of 60 hydatid cysts collected from cattle (n = 40) and buffalo (n = 20), 52 isolates were identified as E. granulosus (s.s.) (G1, G3) and 8 as E. ortleppi (G5). The detection of the G5 genotype represents the first in Pakistan. The phylogeny inferred by the Bayesian method using nucleotide sequences of cox1-nad1 further confirmed their identity. The diversity indices indicated a high haplotype diversity and a low nucleotide diversity. The negative values of Tajima’s D and Fu’s Fs test demonstrated deviation from neutrality suggesting a recent population expansion. Conclusions: To the best of our knowledge, this report described the genetic variation of E. granulosus population for the first time in Pakistan using the complete cox1 and nad1 mitochondrial genes and confirms E. ortleppi as one of the causative agents of CE among livestock in Pakistan. While this report will contribute to baseline information for CE control, more studies considering species diversity and distribution in different hosts across unstudied regions of Pakistan are highly needed.