Dataset Information

Molecular identification of Cryptosporidium spp. in alpacas (Vicugna pacos) in China.

ABSTRACT: Cryptosporidium is a ubiquitous protozoan in human and animals. To investigate the genetic diversity of Cryptosporidium spp. in alpaca (Vicugna pacos) in China, 484 fecal samples from alpacas were collected at nine sites, and Cryptosporidium spp. were screened with PCR amplification of the small subunit ribosome RNA (SSU rRNA) locus. Cryptosporidium spp. infected 2.9% (14/484) of the alpacas. Of the nine collection sites, two were positive for Cryptosporidium, Wensu (3.0%, 3/100) and Qinghe (31.4%, 11/35). Three Cryptosporidium species were identified: C. parvum (n = 2), C. ubiquitum (n = 1), and C. occultus (n = 11). Cryptosporidium parvum and C. ubiquitum were further subtyped with the 60-kDa glycoprotein locus (gp60). The two C. parvum isolates were subtype IIdA15G1, but the one C. ubiquitum isolate was not subtyped successfully. A phylogenetic analysis indicated that the Cryptosporidium isolates clustered with previously identified species. To our knowledge, this is the first report of Cryptosporidium infections in alpacas in China and provides baseline data for the study of Cryptosporidium in alpacas in China.

SUBMITTER: Zhang Q

PROVIDER: S-EPMC7316868 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Molecular identification of Cryptosporidium spp. in alpacas (Vicugna pacos) in China.

Zhang Qiyuan Q Li Junqiang J Li Zhiguo Z Xu Chunyan C Hou Minyu M Qi Meng M

International journal for parasitology. Parasites and wildlife 20200620

Cryptosporidium is a ubiquitous protozoan in human and animals. To investigate the genetic diversity of Cryptosporidium spp. in alpaca (Vicugna pacos) in China, 484 fecal samples from alpacas were collected at nine sites, and Cryptosporidium spp. were screened with PCR amplification of the small subunit ribosome RNA (SSU rRNA) locus. Cryptosporidium spp. infected 2.9% (14/484) of the alpacas. Of the nine collection sites, two were positive for Cryptosporidiu ...[more]

PMID: 32612927

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Project description:BackgroundGastrointestinal nematodes (GINs) can cause significant economic losses in alpacas due to lowered production of fibre and meat. Although no anthelmintics are registered for use in alpacas, various classes of anthelmintics are frequently used to control parasitic gastroenteritis in alpacas in Australia and other countries. Very little is known about the current worm control practices as well as the efficacy of anthelmintics used against common GINs of alpacas. This study aimed to assess the existing worm control practices used by Australian alpaca farmers and to quantify the efficacy of commonly used anthelmintics against GINs of alpacas.MethodsAn online questionnaire survey was conducted to assess current worm control practices on 97 Australian alpaca farms, with an emphasis on the use of anthelmintics. Of this group of 97 alpaca farms, 20 were selected to assess the efficacy of eight anthelmintics and/or their combinations (closantel, fenbendazole ivermectin, monepantel, moxidectin and a combination of levamisole, closantel, albendazole, abamectin) using the faecal egg count reduction test (FECRT). A multiplexed-tandem PCR (MT-PCR) was used to identify the prevalent nematode genera/species.ResultsThe response rate for the questionnaire was 94% (91/97). Almost half of the respondents kept alpacas with sheep and cattle, and 26% of respondents allowed alpacas to co-graze with these ruminants. Although only 63% respondents perceived worms to be an important health concern for alpacas, the majority of respondents (89%) used anthelmintics to control GINs of alpacas. The commonly used anthelmintics were macrocyclic lactones, monepantel, benzimidazoles, levamisole, closantel and their combinations, and they were typically administered at the dose rate recommended for sheep. The FECRT results showed that a combination of levamisole, closantel, albendazole and abamectin was the most effective dewormer followed by single drugs, including monepantel, moxidectin, closantel, fenbendazole and ivermectin. Haemonchus spp. were the most commonly resistant nematodes followed by Trichostrongylus spp., Camelostrongylus mentulatus, Ostertagia ostertagi and Cooperia spp.ConclusionsThis is the first study aimed at assessing worm control practices and efficacy of commonly used anthelmintics in alpacas in Australia. Our findings document the extent of anthelmintics resistance on Australian alpaca farms and identify those anthelmintics that are still effective against GINs of alpacas.

Project description:BACKGROUND:Melanocytes are derived from neural crest stem cells in the embryonic stage. In mature melanocytes, a series of complex enzyme-catalyzed reactions leads to the production of melanins, which determine the hair and skin colors of animals. The process of melanogenesis is complex and can be regulated by mRNA, microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) genes. MiRNAs are a type of endogenous noncoding RNA approximately 22 nt in size that predominantly regulate gene expression by inhibiting translation. miR-380-3p is a candidate miRNA potentially related to melanogenesis. To better understand the mechanism of miR-380-3p melanogenesis regulation, plasmids to overexpress or knockdown miR-380-3p were transfected into alpaca melanocytes, and their effects on melanogenesis were evaluated. RESULTS:In situ hybridization identified a positive miR-380-3p signal in alpaca melanocyte cytoplasm. Luciferase activity assays confirmed that SOX6 is targeted by miR-380-3p. miR-380-3p overexpression and knockdown in alpaca melanocytes respectively downregulated and upregulated SOX6 expression at the mRNA and protein levels. Additionally, miR-380-3p overexpression and knockdown, respectively, in alpaca melanocytes decreased and increased the mRNA levels of melanin transfer-related genes, including microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosine-related protein-1 (TYRP1), and dopachrome tautomerase (DCT). In contrast, miR-380-3p overexpression and knockdown respectively increased and decreased the mRNA levels of β-catenin. Additionally, the effect of miR-380-3p on melanogenesis was assessed by Masson-Fontana melanin staining. CONCLUSIONS:The results demonstrated that miR-380-3p targeted SOX6 to regulate melanogenesis by influencing β-catenin and MITF transcription and translation, which reduced the expression of downstream genes, including TYR, TYRP1, and DCT. These results provide insights into the mechanisms through which miR-380-3p controls melanogenesis.

Project description:BackgroundEukaryotic pathogens, including Cryptosporidium, Giardia and Enterocytozoon, have been implicated in neonatal diarrhoea, leading to marked morbidity and mortality in the alpaca (Vicugna pacos) and llama (Lama glama) around the world. Australia has the largest population of alpacas outside of South America, but very little is known about these pathogens in alpaca populations in this country. Here, we undertook the first molecular epidemiological survey of Cryptosporidium, Giardia and Enterocytozoon in V. pacos in Australia.MethodsA cross-sectional survey of 81 herds, comprising alpacas of 6 weeks to 26 years of age, were sampled from the six Australian states (Queensland, New South Wales, Victoria, South Australia, Tasmania and Western Australia) across the four seasons. PCR-based sequencing was employed, utilising genetic markers in the small subunit of the nuclear ribosomal RNA (SSU) and 60-kilodalton glycoprotein (gp60) genes for Cryptosporidium, triose-phosphate isomerase (tpi) gene for Giardia duodenalis and the internal transcribed spacer region (ITS) for Enterocytozoon bieneusi.ResultsPCR-based analyses of 81 faecal DNA samples representing 1421 alpaca individuals detected Cryptosporidium, Giardia and/or Enterocytozoon on 15 farms in New South Wales, Victoria and South Australia, equating to 18.5% of all samples/herds tested. Cryptosporidium was detected on three (3.7%) farms, G. duodenalis on six (7.4%) and E. bieneusi on eight (9.9%) in two or all of these three states, but not in Queensland, Tasmania or Western Australia. Molecular analyses of selected faecal DNA samples from individual alpacas for Cryptosporidium, Giardia and/or Enterocytozoon consistently showed that alpacas of ≤ 6 months of age harboured these pathogens.ConclusionsThis first molecular investigation of Cryptosporidium, Giardia and Enterocytozoon in alpaca subpopulations in Australia has identified species and genotypes that are of likely importance as primary pathogens of alpacas, particularly young crias, and some genotypes with zoonotic potential. Although the prevalence established here in the alpaca subpopulations studied is low, the present findings suggest that crias are likely reservoirs of infections to susceptible alpacas and/or humans. Future studies should focus on investigating pre-weaned and post-weaned crias, and on exploring transmission patterns to establish what role particular genotypes play in neonatal or perinatal diarrhoea in alpacas and in zoonotic diseases in different states of Australia.

Dataset Information

Molecular identification of Cryptosporidium spp. in alpacas (Vicugna pacos) in China.

Publications

Molecular identification of <i>Cryptosporidium</i> spp. in alpacas (<i>Vicugna pacos</i>) in China.

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