First cross-sectional, molecular epidemiological survey of Cryptosporidium, Giardia and Enterocytozoon in alpaca (Vicugna pacos) in Australia

Abstract

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.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-3055-6) contains supplementary material, which is available to authorized users.

Molecular detection of Enterocytozoon bieneusi in alpacas (Vicugna pacos) in Xinjiang, China

“…Three out of the five alpaca herds were positive, and the highest infection rate (42.9%, 15/35) was detected in animals from Qinghe (Table 1). The overall infection rate of E. bieneusi (15.1%) in this study was lower than that reported in alpacas in the highlands of Peru (51.6%) [3], and higher than that in Australia (9.9%) [4]. Currently, E. bieneusi has been isolated from humans, nonhuman primates, pigs, cattle, sheep, yaks, deer, cats and dogs, chickens, rodents, and snakes, as well as urban wastewater in China [18].…”

“…Among the three herds that were positive for E. bieneusi , genotype ALP3 ( n = 15) was detected in Qinghe, genotype ALP8 ( n = 1) in Hejing, and genotypes ALP3, ALP7, and P in Wensu (Table 1). Until now, molecular investigation of E. bieneusi in alpacas has been limited to two studies from farmed alpacas in Peru and Australia [3, 4], and two studies from captive alpacas in Chinese zoos [5, 6]. A total of 14 E. bieneusi genotypes (P, ALP1–8, Type IV, D, BEB6, J, and CHALTI) have been identified in alpacas worldwide [3–6], and genotype ALP1 was dominant in Peru and Australia [3, 4] (Table 2).…”

“…Until now, molecular investigation of E. bieneusi in alpacas has been limited to two studies from farmed alpacas in Peru and Australia [3, 4], and two studies from captive alpacas in Chinese zoos [5, 6]. A total of 14 E. bieneusi genotypes (P, ALP1–8, Type IV, D, BEB6, J, and CHALTI) have been identified in alpacas worldwide [3–6], and genotype ALP1 was dominant in Peru and Australia [3, 4] (Table 2). Interestingly, only two known genotypes (ALP3 and P) and two novel genotypes (ALP7 and ALP8) were identified in this study, with genotype ALP3 (57.1%, 16/28) being the most frequent genotype, followed by genotype P (42.8%, 9/28).…”

“…Interestingly, only two known genotypes (ALP3 and P) and two novel genotypes (ALP7 and ALP8) were identified in this study, with genotype ALP3 (57.1%, 16/28) being the most frequent genotype, followed by genotype P (42.8%, 9/28). Genotype ALP3 was only detected in two farmed alpaca samples in Peru and Australia [3, 4], whereas genotype P was first identified in captive llamas ( Lama glama ) from Munich Zoo [1], and then later detected in fecal samples of alpacas from Peru and Australia [3, 4]. The differences in the predominance of E. bieneusi genotypes in different areas indicate that E. bieneusi infection in alpacas may exhibit regional differences.…”

“…positive (%)Genotype (no. )ReferenceChinaZoo43 (75.0)CHALT1 (1), J (2)[5]ChinaZoo11 (100)BEB6 (1)[6]PeruFarms12665 (51.6)ALP1 (48), ALP2 (1), ALP3 (1), ALP4 (1), ALP5 (1), ALP6 (1), BEB6 (1), D (2), P (5), Type IV (4) [3]AustraliaFarms818 (9.9)ALP1 (5), ALP3 (1), P (2)[4]ChinaFarms18528 (15.1)ALP3 (16), ALP7 (2), ALP8 (1), P (9) This studyTotalALP1 (53), ALP2 (1), ALP3 (18), ALP4 (1), ALP5 (1), ALP6 (1), ALP7 (2), ALP8 (1), BEB6 (2), CHALT1 (1), D (2), J (2), P (16), Type IV (4)…”