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Background: Paratuberculosis, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic granulomatous enteritis that affects domestic and wild ruminants and camelids. The disease has rarely been reported in alpacas in Germany. This publication describes epidemiologically independent cases of paratuberculosis in two alpacas in Germany., Case Presentation: Two alpacas, a 26-year-old female zoo animal (case 1) and a 2.5-year-old breeding stallion from a private owner (case 2), presented with progressive emaciation, leading to death (case 2) or euthanasia (case 1) because of deteriorating general condition. In both cases typical granulomatous lesions in the intestinal mucosa and mesenteric lymph nodes were found. In case 2, other lymph nodes were severely enlarged and MAP was detected in the mandibular lymph node, lung, and liver by qPCR. The MAP isolates differed between the alpacas, with two distinct phylogenetic clades (Clade 1 and 8) within Subgroup A of the MAP-C type group and two distinct INMV profiles (INMV 2 and 1) found. These genotypes have been identified in cattle and goats in different regions in Germany. The genotype isolated from case 1 has been detected in goats from the zoo since 2011, indicating transmission between these species., Conclusions: MAP can cause severe clinical disease in alpacas of variable age and under different husbandry conditions. Therefore, paratuberculosis should be considered for differential diagnosis in alpacas with emaciation and poor general condition. Although not definitely shown, cross-species infection between ruminant species and camelids is exceedingly likely., Competing Interests: Declarations. Ethics approval and consent to participate: An Animal Ethics approval was waived, because the data of both cases originated from routine diagnostics and follow up of clinical disease of animals. All diagnostic and laboratory methods were carried out in accordance with relevant guidelines and regulations. The National Reference Laboratories of the FLI have been accredited by the Deutsche Akkreditierungsstelle GmbH (DAkkS) according to ISO/IEC 17025. The authors obtained informed consent of both owners for the use of the animals and the data collected from them. Human ethics: not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Anemia is a common problem in South American camelids (SACs). Infections with Candidatus Mycoplasma haemolamae (CMh), a cell-wall free, hemotropic bacterium, are often suspected to be an important cause of anemia, as the pathogen infects the erythrocytes and is found in the blood of up to 30% of SACs. The information on the clinical signs of animals infected with this pathogen vary widely. Most infections are clinically inapparent. Treatment is usually carried out with oxytetracycline. A detailed overview of the clinical and hematological findings in 13 alpacas infected with Candidatus M. haemolamae (CMh+), based on patients from our university clinic and comparing those findings with the results of 22 negative alpacas (CMh-) is provided. Assignment to both groups was based on the PCR result. No relevant clinical or hematological differences between CMh+ and CMh- were found, the clinical signs in CMh+ were usually due to comorbidities. The examination of a blood smear alone proved to be insufficient; a PCR test should be carried out to confirm or rule out an infection. A critical review of the need for antibiotic treatment on the basis of a positive test result alone is recommended., (© 2024. The Author(s).)

Two strictly anaerobic, Gram-stain-negative rod-shaped bacterial isolates, A2-P53 T and A1-P5, were isolated from an enrichment of fecal material from two alpacas (Vicugna pacos). Based on a comparative 16S rRNA gene sequence analysis, the isolates were assigned to the genus Bacteroides with the highest sequence similarities to Bacteroides koreensis YS-aM39T (A2- P53T 97.7 % and A1-P5 97.9 %). Additionally, the average nucleotide identity and digital DNA-DNA hybridization values between these isolates and their closest relatives within Bacteroides were less than 92.1 % and 49.1 %, respectively. The average nucleotide identity between isolates A2-P53 T and A1-P5 was 99.9 %. The predominant cellular fatty acid for isolates A2-P53 T and A1-P5 was C 15:0 antesio. The G+C % content of the isolates was 41.7 %. Based on biochemical, phylogenetic, genotypic, and chemotaxonomic criteria, these isolates A2-P53 T and A1-P5 represent two individual strains of a novel species within the genus Bacteroides for which the name Bacteroides vicugnae sp. nov. is proposed. The type strain of this species is strain A2-P53 T (CCUG 77273 T  = CCM 9377 T  = NRRL B-65693 T )., Competing Interests: Declaration of competing interest The authors declare that they have no indirect or direct conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A strictly anaerobic, Gram-stain-negative rod-shaped bacterium, designated A1-XYC3 T , was isolated from the faeces of an alpaca ( Lama pacos ). On the basis of the results of a comparative 16S rRNA gene sequence analysis, the isolate was assigned to the genus Clostridium with the highest sequence similarities to Clostridium magnum DSM 2767 T (96.8 %), Clostridium carboxidivorans P7 T (96.3 %) and Clostridium aciditolerans JW/YJL-B3 T (96.1 %). The average nucleotide identity between A1-XYC3 T , C. magnum, C. carboxidivorans and C . aciditolerans was 77.4, 76.1 and 76.6  %, respectively. The predominant components of the cellular fatty acids of A1-XYC3 T were C 14 : 0 , C 16 : 0 and summed feature 10, containing C 18:0 /C 17:0 cyclo. The DNA G+C content was 32.4 mol%. On the basis of biochemical, phylogenetic, genotypic and chemotaxonomic criteria, this isolate represents a novel species within Clostridium sensu stricto for which the name Clostridium tanneri sp. nov. is proposed. The type strain of this species is strain A1-XYC3 T (=CCM 9376 T =NRRL B-65691 T ).

Hemotrophic mycoplasmas (hemoplasmas) are epierythrocytic bacteria that infect wild and domestic animals, and can cause anemia in some of them. They are considered emerging and zoonotic pathogens, causing serious health problems in wildlife. Candidatus Mycoplasma haemolamae is the only species of hemoplasma that infects domestic South American camelids (alpacas and llamas), with limited studies in wild camelids. Therefore, the objective of this study was to determine the prevalence of Candidatus M. haemolamae in vicunas (Vicugna vicugna) from the Pampa Galeras National Reserve, located in the Ayacucho region of Peru, using molecular diagnosis. For this, blood samples from 79 vicunas were collected, which were molecularly analyzed by partially amplifying the 16S ribosomal RNA gene of Mycoplasma sp. Fourteen vicunas (17.7 %) were positive for the molecular diagnosis of Mycoplasma sp. All PCR-positive products were sequenced and showed more than 99 % identity with Candidatus M. haemolamae. Statistical analysis showed that tick-infested vicunas had 6.10 odds of presenting Candidatus M. haemolamae compared with tick-free vicunas. Sex and age were not associated with Candidatus M. haemolamae infections. This is the first report of hemoplasmas in vicunas, a wild South American camelid, demonstrating that the pathogen can have both a domestic and a wild life cycle. Future studies are necessary to know the current situation of this pathogen in domestic and wild camelids from other locations in Peru., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study aimed to perform a molecular survey and identification of hemotropic Mycoplasma spp. in domestic South American Camelids from Southern Chile. Conventional PCR (cPCR) for hemotropic Mycoplasma spp. based on 16S rRNA gene (620bp fragment) was performed in 87 EDTA-blood samples taken from 48 llamas (Lama glama) and 39 and alpacas (Vicugna pacos) from to Temuco, La Araucanía region and Valdivia, Los Rios region, Southern Chile. 16S rRNA hemotropic Mycoplasma PCR-positive were sequenced for species identification, phylogenetic and haplotype analyses, and further tested by cPCR targeting a fragment (160-210 bp) of the RNaseP (rnpB) gene. Based upon 16S rRNA cPCR results, the overall hemotropic Mycoplasma spp. occurrence in Southern camelids was 9.2% (8/87 [95% CI (4.0-17.3%)]), with five positive alpacas (12.8%; 5/39 [95% CI (4.3-27.4%)]) and three llamas (6.3%; 3/48 [95% CI (1.7-17.2%)]). All 16S rRNA PCR-positive samples were negative for the rnpB gene. Obtained 16S sequences presented high identity (99-100%) by BLASTn analysis to 'Candidatus Mycoplasma haemolamae' from an alpaca in the United Kingdom. Phylogenetic and haplotype analyses of the 16s rRNA gene showed high similarity among 'Candidatus M. haemolamae' sequences of this study and the ones from North America, Europe, and Asia evidencing a low diversity of Chilean samples, with only one haplotype detected (#1). Haplotype #1 from South American Camelids in Chile was worldwide distributed and observed in North America, Europe, and Asia. 'Candidatus M. haemolamae' detected for the first time in South American camelids in Southern Chile had low diversity and was worldwide spread., (Copyright © 2021. Published by Elsevier B.V.)

Four novel independent strains of Streptococcus spp. were isolated from faeces of alpaca (SL1232 T ), cattle (KCJ4950), and from respiratory tract of wild California sea lions (CSL7508 T , CSL7591 T ). The strains were indole-, oxidase- and catalase-negative, non-spore-forming, non-motile Gram-positive cocci in short and long chains, facultative anaerobes. The 16S rRNA gene of SL1232 T and KCJ4950 shared 99.40-99.60% nucleotide similarity to strains of S. equinus, S. lutetiensis, S. infantarius , and the 16S rRNA gene of CSL7508 T and CSL7591 T demonstrated 98.72 and 98.92% similarity, respectively, to S. marimammalium . All other known Streptococcus species had the 16S rRNA gene sequence similarities of ≤95%. The genomes were sequenced for the novel strains. Average nucleotide identity (ANI) analysis for strains SL1232 T and KCJ4950, showed the highest similarity to S. equinus, S. lutetiensis, and S. infantarius with 85.21, 87.17, 88.47, 85.54, 87.47 and 88.89%, respectively, and strains CSL7508 T and CSL7591 T to S. marimammalium with 87.16 and 83.97%, respectively. Results of ANI were confirmed by pairwise digital DNA-DNA hybridization and phylogeny, which also revealed that the strains belong to three novel species of the genus Streptococcus . Phenotypical features of the novel species were in congruence with closely related members of the genus Streptococcus and gave negative reactions with the tested Lancefield serological groups (A-D, F and G). MALDI-TOF mass spectrometry supported identification of the species. Based on these data, we propose three novel species of the genus Streptococcus , for which the name Streptococcus vicugnae sp. nov. is proposed with the type strain SL1232 T (=NCTC 14341 T =DSM 110741 T =CCUG 74371 T ), Streptococcus zalophi sp. nov. is proposed with the type strain CSL7508 T (=NCTC 14410 T =DSM 110742 T =CCUG 74374 T ) and Streptococcus pacificus sp. nov. is proposed with the type strain CSL7591 T (=NCTC 14455 T =DSM 111148 T =CCUG 74655 T ). The genome G+C content is 36.89, 34.85, and 35.34 % and draft genome sizes are 1906993, 1581094 and 1656080 bp for strains SL1232 T , CSL7508 T , and CSL7591 T , respectively.

Information about livestock carrying methicillin-resistant coagulase-negative staphylococci and mammaliicocci (MRCoNS/MRM) is scarce. The study was designed to gain knowledge of the prevalence, the phenotypic and genotypic antimicrobial resistance and the genetic diversity of MRCoNS/MRM originating from ruminants and New World camelids. In addition, a multi-locus sequence typing scheme for the characterization of Mammaliicoccus (formerly Staphylococcus) sciuri was developed. The study was conducted from April 2014 to January 2017 at the University Clinic for Ruminants and the Institute of Microbiology at the University of Veterinary Medicine Vienna. Seven hundred twenty-three nasal swabs originating from ruminants and New World camelids with and without clinical signs were examined. After isolation, MRCoNS/MRM were identified by MALDI-TOF, rpoB sequencing and typed by DNA microarray-based analysis and PCR. Antimicrobial susceptibility testing was conducted by agar disk diffusion. From all 723 nasal swabs, 189 MRCoNS/MRM were obtained. Members of the Mammaliicoccus (M.) sciuri group were predominant (M. sciuri (n = 130), followed by M. lentus (n = 43), M. fleurettii (n = 11)). In total, 158 out of 189 isolates showed phenotypically a multi-resistance profile. A seven-loci multi-locus sequence typing scheme for M. sciuri was developed. The scheme includes the analysis of internal segments of the house-keeping genes ack, aroE, ftsZ, glpK, gmk, pta1 and tpiA. In total, 28 different sequence types (STs) were identified among 92 selected M. sciuri isolates. ST1 was the most prevalent ST (n = 35), followed by ST 2 (n = 15), ST3 and ST5 (each n = 5), ST4 (n = 3), ST6, ST7, ST8, ST9, ST10 and ST11 (each n = 2)., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

During the last few years, the number of cases of Trichophyton verrucosum isolation from humans suffering from mycoses has been constantly increasing, which is correlated with the presence of an increasing number of outdoor breeding farms. Farmers and their families as well as veterinarians and technicians involved in handling the animals are at a higher risk of infection. One of the most important aims of mycological diagnostics is epidemiological analysis. Typically, the history of the disease is not sufficient to indicate reliably and eliminate the outbreak of infection. PCR fingerprinting methods are a useful tool in this type of analysis, which is presented in this study. The main aim is to present diagnostic and epidemiological analyses of dermatophyte isolates from llamas and their breeder. In two llamas, round alopecia sites or ca. 2-cm excoriations covered with thickened scaling epidermis were noticed at the border of the head and neck with a distinct tendency towards hair loss. Tinea unguium was noticed in a nail of the breeder's right hand. Direct analysis of the material from the clinical lesions revealed the presence of arthrospores. The macro- and micromorphology of the isolates were homogeneous and characteristic for T. verrucosum. The identification analysis based on the ITS sequences confirmed the previous morphological diagnostic examination. The MP-PCR and MSP-PCR analysis indicated high invariability of the genomes of the strains isolated from the human and animals. The epidemiological research has indicated an identical source of dermatophyte infection in the breeder and the lamas. To sum up, the number of pets and farm animals is increasing and dermatologists should always be informed about possible dermatophyte transmission sources. The possibility of transmission of zoophilic dermatophytes from humans to animals is a suggestion for further analysis; therefore, this type of transmission should be considered in dermatological studies., (© 2019 Blackwell Verlag GmbH.)

Ten strains of an Actinobacillus -like organism were isolated from alpaca ( Vicugna pacos ) in the UK over a period of 5 years, with no known epidemiological linkages. The isolates are distinct, based on both phenotype and genotype, from any previously described Actinobacillus species. Molecular analysis, based on 16S rRNA, rpoB and infB gene sequences, placed the isolates as a novel, early branching, lineage within the currently recognised Actinobacillus sensu stricto . In agreement with the results of the single-gene analysis, average nucleotide identity values, based on whole genome sequences, showed very similar identities to a number of members of the Actinobacillus sensu stricto notably Actinobacillus equuli , Actinobacillus suis and Actinobacillus ureae . At least two phenotypic characteristics differentiate the alpaca isolates from other Actinobacillus sensu stricto species, and from taxa likely falling within this group but awaiting formal species description, with Actinobacillus anseriformium and A. equuli subsp. haemolyticus being the most closely related phenotypically. The alpaca isolates can be differentiated from A. anseriformium by production of β-galactosidase (ONPG) and acid from raffinose, and from A. equuli subsp. haemolyticus by production of acid from d-sorbitol and failure to produce acid from d-xylose. Isolates were obtained from multiple sites in alpaca including respiratory tract, alimentary tract and internal organs although further evidence is required to understand any pathogenic significance. Based on the results of characterization described here, it is proposed that the isolates constitute a novel species, Actinobacillus vicugnae sp. nov. The type strain is W1618 T (LMG30745 T NCTC14090 T ) isolated in the UK in 2012 from oesophageal ulceration in an alpaca ( Vicugna pacos ).

Background: Mycoplasma haemolamae (Mhl) and gastrointestinal nematodes can cause anemia in camelids. Control programs aim to suppress parasitism without promoting anthelminthic resistance, but few evidence-based guidelines define acceptable parasite loads in camelids., Hypothesis/objectives: In clinically healthy nonanemic camelids, compare erythrocyte variables to Mhl real-time PCR status and to fecal egg count (FEC). Determine the FEC threshold above which erythrocyte variables are consistently below reference interval medians., Animals: One hundred fourteen client-owned adult alpacas and llamas., Methods: In a cross-sectional study, whole blood in ethylenediaminetetraacetic acid (EDTA) was assessed for packed cell volume (PCV) by centrifugation, erythrocyte count (RBC), and hemoglobin concentration (HGB) using an ADVIA120 analyzer, and Mhl using real-time PCR. Trichostrongyle eggs per gram (epg) were counted by modified McMaster test on freshly collected feces. Significant differences in erythrocyte variables based on Mhl status and FEC thresholds were assessed by independent t test and one-way ANOVA, respectively., Results: Packed cell volume, RBC, and HGB were not significantly different between Mhl-positive and Mhl-negative animals, but were significantly lower in animals with FEC >1000 epg compared to those with <500 epg. All animals with FEC >600 epg had RBC and HGB below the reference interval median. All animals with FEC >750 epg had PCV below the reference interval median., Conclusions and Clinical Importance: In healthy nonanemic camelids, positive Mhl PCR is not associated with lower erythrocyte variables and such animals may not warrant treatment. Fecal egg count >600-750 epg has a negative effect on erythrocyte variables, and may be a guide for deworming protocols., (© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

Enterocytozoon bieneusi, an obligate intracellular pathogen, can infect a wide variety of hosts. This study aimed to determine the prevalence and molecular characteristics of E. bieneusi in alpacas (Vicugna pacos) in China. A total of 185 alpaca fecal samples were collected from five herds in Tacheng, Wensu, Hejing, Qinghe, and Nilka counties in Xinjiang Uygur Autonomous Region. Enterocytozoon bieneusi was detected by nested PCR of the internal transcribed spacer (ITS) region. Twenty-eight fecal samples (15.1%, 28/185) were positive for E. bieneusi, with the highest prevalence in alpacas from Qinghe (42.9%, 15/35). Four E. bieneusi genotypes were identified, which included two known (P and ALP3) and two novel (ALP7 and ALP8) genotypes. Genotype ALP3 was the dominant genotype (57.1%, 16/28), followed by genotypes P (32.1%, 9/28), ALP7 (7.1%, 2/28), and ALP8 (2.6%, 1/28). Phylogenetic analysis revealed that three genotypes (P, ALP7, and ALP3) clustered into group 1, whereas genotype ALP8 clustered into group 8. This is the first report of E. bieneusi infection and genetic diversity in alpacas from Xinjiang, China., (© Q. Zhang et al., published by EDP Sciences, 2019.)

Background: Streptococcus agalactiae is pathogenic for both animals and humans. In dairy cattle it commonly causes mastitis, with great economic losses, and there is scientific evidence of mastitis, caseous lymphadenitis, contagious skin necrosis and purulent infections associated with S. agalactiae in camels (Camelus dromedarius) as well. In humans, it is a common component of the respiratory and gastrointestinal microflora, but it can also act as a pathogen, especially in elderly people and immunocompromised patients, as well as in pregrant women and newborns., Case Presentation: A 10-year old non-pregnant female llama (Lama glama) was conferred to the Institute for Animal Health Control, in Bolzano for necropsy after sudden death. The animal had not shown unusual behaviour and had a low to normal nutritional condition (body condition score 2/5). The breeder had reported a chronic suppurative subcutaneous infection in the intermandibular area, resistant to therapy (therapy unknown). After necropsy, several samples were processed for histological, bacteriological and parasitological examinations., Conclusions: This report describes, to the best of our knowledge, the first isolation of S. agalactiae in llamas (Lama glama). The animal came from a herd that counts approximately 200 South American camelids (llamas, alpacas) along with several horses, chicken, rabbits, cats and dogs; this farm offers services, such as trekking and pet therapy activities.

Background: Eukaryotic 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., Methods: A 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., Results: PCR-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., Conclusions: This 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.

The aim of this study was to determine the prevalence, the antimicrobial resistance patterns and the genetic diversity of methicillin-resistant Staphylococcus aureus (MRSA) from Austrian ruminants and New World camelids that were treated at the University of Veterinary Medicine, Vienna. Between April 2014 and January 2017, 723 nasal swabs originating from ruminants and New World camelids were examined. MRSA isolates were characterized by mecA/mecA1/mecC PCRs and by DNA microarray analysis. They were genotyped by spa typing, dru typing, MLST and MLVA. Glycopolymer fingerprinting by FTIR spectroscopy was also performed. Antimicrobial susceptibility testing was conducted by agar disk diffusion. Twelve MRSA isolates were mecA-positive, whereas three were mecC-positive. The MRSA isolates carried five different SCCmec elements, and belonged to three sequence types (ST45, ST130, ST398). The MRSA isolates displayed seven different resistance phenotypes. The present study describes for the first time mecC-carrying MRSA isolates originating from domesticated animals in Austria. More systematic studies are needed to unravel the role of ruminants and New World camelids as reservoirs for MRSA as a potential risk for zooanthropogenic transmission., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Aim: To determine the prevalence of infection with Candidatus Mycoplasma haemolamae (Mhl), antibodies to bovine viral diarrhoea virus (BVDV), and BVDV antigen, and the prevalence of animals with elevated faecal nematode egg counts (FEC) in a sample of adult New Zealand alpaca (Vicugna pacos)., Methods: Blood samples were obtained from 175 alpaca, collected from 15 farms around New Zealand, and from 31 samples sent to a diagnostic laboratory for routine haematology. Blood smears (n=170) were examined microscopically for the presence of haemoplasma, and DNA was extracted from whole blood (n=206) for real-time PCR testing for Mhl. Packed cell volume (PCV) was determined for 193 samples. Serum samples (n=195) were tested for BVDV antibody using ELISA, and for BVDV antigen using a real-time PCR assay. Faecal samples were collected from 143 animals; FEC were measured, and samples pooled for larval culture., Results: No haemoplasma organisms were present on blood smear examination. Of the 206 blood samples, two (from the same farm) were positive for Mhl by real-time PCR testing, giving a prevalence of infection with Mhl of 0.97%. Of the 195 serum samples tested, four (2.1%) were positive for antibodies to BVDV; animals with BVDV antibodies were from 3/15 (20%) farms, none of which farmed cattle. None of the serum samples were positive by PCR for BVDV antigen. The median FEC was 50 epg (min 0, max 4,700), with 55/143 (38.5%) samples having 0 epg, and 33/143 (23.1%) having ≥250 epg. Haemonchus spp. were the most common nematodes present in faecal larval cultures from the North Island. Log 10 FEC was negatively associated with PCV (p=0.02), and was higher in males than females (p<0.001), and in animals that were positive compared with negative for Mhl (p=0.022)., Conclusions and Clinical Relevance: The number of alpaca infected with Mhl was low, as was the seroprevalence of BVDV. Gastrointestinal parasitism was, however, a common finding in this sample of New Zealand alpaca.

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis in domestic ruminants and New World Camelids (NWC). Hepatitis E virus (HEV) is an important public health concern worldwide. The virus has been identified in several species, some of them serving as a reservoir for zoonotic HEV strains. Husbandry and breeding of llamas and alpacas have increased in Austria in recent years. Therefore, the aim of the present study was to evaluate the prevalence of MAP and HEV in NWC in Austria. Altogether 445 animals, originating from 78 farms were enrolled in the study. Of the animals sampled, 184 (41.35%) were llamas and 261 (58.65%) were alpacas. 443 blood samples for MAP-ELISA and 399 faecal samples for quantitative PCR (qPCR) and culture for MAP as well as for HEV detection by RT-qPCR have been collected. All of the 399 animals tested for shedding of MAP were negative by faecal solid culture. Using qPCR, 15 (3.8%) of the animals were MAP positive and 384 (96.2%) negative. Out of the 443 serum samples examined for specific antibodies against MAP by ELISA, 6 (1.4%) were positive, 1 (0.2%) was questionable and 436 (98.4%) samples were negative. All faecal samples were tested negative for HEV.

Objective: To describe the bacterial flora of the normal conjunctiva of Huacaya alpacas (Vicugna pacos) and to determine the effect of age and gender on this flora., Animals Studied: Fifty Huacaya alpacas., Procedures: After a complete ophthalmic examination, conjunctival swabs were obtained from both eyes and cultured for aerobic and anaerobic bacteria. Logistic and Poisson regression analyses were used to evaluate the effect of age and gender on bacterial isolation., Results: Four animals were excluded because of signs of external ocular disease. Of the remaining 46 alpacas, bacteria were recovered from 96.7% (89/92) of the eyes. A total of 190 bacterial isolates were cultured with a mean of 2.1 bacterial isolates per eye. The majority of isolates (70%) were Gram-positive. Staphylococcus xylosus (44/190: 23.2%) predominated, followed by viridans streptococci (32/190: 16.8%) and Pantoea agglomerans (24/190: 12.6%). Other frequently isolated bacteria included Rothia mucilaginosa (12/190: 6.3%), Staphylococcus equorum (12/190: 6.3%), Bacillus species (9/190: 4.7%), Moraxella ovis (9/190: 4.7%), and Moraxella catarrhalis (6/190: 3.2%). Statistical analysis showed that alpacas harboring viridans streptococci and Moraxella species were significantly younger. Gender did not significantly affect type of bacterial isolation. There appeared to be no significant effect of age or gender on number of bacteria isolated., Conclusions: Gram-positive aerobes were most commonly cultured, with S. xylosus and viridans streptococci predominating. To the best of the authors' knowledge, this is the first report describing the presence of Moraxella species in the healthy conjunctival sac of alpacas. Alpacas harboring viridans streptococci and Moraxella species were significantly younger., (© 2015 American College of Veterinary Ophthalmologists.)

Two novel actinobacteria, designated strains YIM 101617(T) and YIM 101632(T), were isolated from Lama pacos (alpaca) and Viverra zibetha (civet) faeces in Yunnan Wild Animal Park in Yunnan province, southwestern China. Both strains should be placed in genus Enteractinococcus based on phylogenetic analysis. Based on 16S rRNA gene sequence analysis, strain YIM 101617(T) exhibits high similarity to Enteractinococcus fodinae DSM 22966(T) (97.70 %) and Enteractinococcus coprophilus YIM 100590(T) (97.45 %), whilst YIM 101632(T) exhibits high similarity to Enteractinococcus coprophilus YIM 100590(T) (97.25 %), and the similarity between YIM 101617(T) and YIM 101632(T) is 95.90 %. However, DNA-DNA hybridization values of the two strains with the type strains in the genus Enteractinococcus were low (<70 %). Most morphological and chemotaxonomic characteristics of the two strains were found to be similar to those of species in the genus Enteractinococcus but also some differences were observed. The DNA G+C contents of strains YIM 101617(T) and YIM 101632(T) were determined to be 55.9 and 56.4 mol%, respectively. Based on these data, the two strains are concluded to represent two different novel species in the genus Enteractinococcus. The names Enteractinococcus lamae sp. nov. (type strain YIM 101617(T)=DSM 27612(T)=CCTCC AB 2013230(T)) and Enteractinococcus viverrae sp. nov. (type strain YIM 101632(T)=KCTC 39552(T)=CCTCC AB 2013280(T)) are proposed, respectively.

Objective: Bovine viral diarrhoea virus (BVDV) and Neospora caninum may cause clinical disease in alpacas. Both diseases are present in the Australian cattle population. The objective of this study was to perform a serological prevalence survey for BVDV and N. caninum exposure in a regional alpaca population of South Australia., Methods: Serum samples were taken from 182 alpacas on 10 farms, which had a combined population of 1308 alpacas. Serological analysis for BVD antibodies was performed using a competitive BVDV antibody ELISA kit. Serological analysis for N. caninum was performed using an anti-Neospora ELISA with a protein G conjugate., Results: Of the 182 alpacas sampled, 5 animals located on three properties were positive for BVDV antibodies, constituting a prevalence of 2.7% (95% confidence interval 1-6%). All samples tested negative for N. caninum antibodies., Conclusion: There is a low BVDV seroprevalence and N. caninum is currently either absent or present at a very low prevalence in this population of alpacas in South Australia. There is serological evidence for the presence of both organisms in South Australian beef and dairy cattle herds. Appropriate biosecurity protocols to minimise the risk of introduction and exposure should be a high priority to maintain this favourable status., (© 2015 Australian Veterinary Association.)

Post-weaning diarrhea and edema disease caused by F18 fimbriated E. coli are important diseases in newly weaned piglets and lead to severe production losses in farming industry. Protective treatments against these infections have thus far limited efficacy. In this study we generated nanobodies directed against the lectin domain of the F18 fimbrial adhesin FedF and showed in an in vitro adherence assay that four unique nanobodies inhibit the attachment of F18 fimbriated E. coli bacteria to piglet enterocytes. Crystallization of the FedF lectin domain with the most potent inhibitory nanobodies revealed their mechanism of action. These either competed with the binding of the blood group antigen receptor on the FedF surface or induced a conformational change in which the CDR3 region of the nanobody displaces the D″-E loop adjacent to the binding site. This D″-E loop was previously shown to be required for the interaction between F18 fimbriated bacteria and blood group antigen receptors in a membrane context. This work demonstrates the feasibility of inhibiting the attachment of fimbriated pathogens by employing nanobodies directed against the adhesin domain.

Nosocomial salmonellosis continues to pose an important threat to veterinary medical teaching hospitals. The objectives of this study were to describe an outbreak of salmonellosis caused by Salmonella enterica serovar Oranienburg within our hospital and to highlight its unique features, which can be used to help mitigate or prevent nosocomial outbreaks in the future. We retrospectively analyzed data from patients that were fecal culture-positive for Salmonella Oranienburg between January 1, 2006, and June 1, 2011, including historical, clinical, and pulsed-field gel electrophoresis (PFGE) data. Salmonella Oranienburg was identified in 20 horses, five alpacas, and three cows during this time frame, with dates of admission spanning the period from August, 2006, through January, 2008. We consider most of these patients to have become infected through either nosocomial or on-farm transmission, as evidenced by molecular subtyping results and supportive epidemiologic data. Interpretation of PFGE results in this outbreak was challenging because of the identification of several closely related Salmonella Oranienburg subtypes. Furthermore, a high percentage of cases were fecal culture-positive for Salmonella Oranienburg within 24 h of admission. These patients initially appeared to represent new introductions of Salmonella into the hospital, but closer inspection of their medical records revealed epidemiologic links to the hospital following the index case. Cessation of this outbreak was observed following efforts to further heighten biosecurity efforts, with no known cases or positive environmental samples after January, 2008. This study demonstrates that a Salmonella-positive culture result within 24 h of admission does not exclude the hospital as the source of infection, and it underscores the important role played by veterinary medical teaching hospitals as nodes of Salmonella infection that can promote transmission outside of the hospital setting.

Background: The prevalence of Johne's disease in alpacas in the United States is unknown. The limits of polymerase chain reaction (PCR) detection of Mycobacterium avium subsp. paratuberculosis (MAP) in alpaca feces have not been determined., Objectives: To evaluate the use of PCR for MAP detection in alpaca feces; and to estimate the prevalence of MAP fecal shedding in alpacas presented to veterinary teaching hospitals., Animals: Alpacas presenting to 4 US veterinary teaching hospitals from November 2009 to February 2011., Methods: Prospective study. Ten dilutions of a wild MAP strain were added to negative alpaca feces and processed for MAP detection by means of a commercial real-time PCR (RT-PCR) assay, and cultured on Herrold's Egg Yolk Medium (HEYM) and liquid broth. The limits of detection for each method were determined. Fecal samples from alpacas admitted to the veterinary teaching hospitals during the study period were evaluated for MAP via PCR and HEYM., Results: The lowest MAP dilution detectable via PCR was 243 MAP colony-forming units (CFU)/g of feces, at which concentration MAP growth was detectable on HEYM. Ten (6%; 95% confidence interval: 3-9%) of the 180 fecal samples collected were positive on PCR., Conclusions and Clinical Importance: Polymerase chain reaction can provide an accurate and rapid detection of MAP fecal shedding in alpacas; and the prevalence of MAP fecal shedding in hospitalized alpacas in 4 US veterinary teaching hospitals was 6%., (Copyright © 2013 by the American College of Veterinary Internal Medicine.)

The high fibrolytic activity and large biomass of strictly-anaerobic bacteria that inhabit the rumen makes them primarily responsible for the degradation of the forage consumed by ruminants. Llamas feed mainly on low quality fibrous roughages that are digested by an active and diverse microflora. The products of this fermentation are volatile fatty acids and microbial biomass, which will be used by the animals. The aim of this study was to detect the three major fiber-digesting anaerobic bacteria in the forestomach contents of llamas by PCR. In this study, we detected Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in the forestomach contents of eight native llamas from Argentina.

Background: An ulcerative dermopathy caused by Mycobacterium ulcerans is described in two alpacas (Vicugna pacos) domiciled in endemic areas of Victoria, Australia., Results: The diagnosis was confirmed in both cases by PCR targeting the M. ulcerans-specific insertion sequence, IS2404. Extensive wound debridement and bandaging was effective in controlling local disease in one alpaca, although the animal was eventually euthanased because of suspected disease recurrence at other anatomical sites. Treatment was not undertaken in the second animal, but the results of a complete necropsy are described. Investigation of the environs of the second animal yielded low levels of M. ulcerans DNA associated with a variety of samples. The potential use of adjunctive antibiotic therapies directed against M. ulcerans infection in this species is discussed., Conclusion: Mycobacterium ulcerans infection should be suspected in alpacas domiciled in endemic areas and presented with ulcerative skin disease., (© 2013 The Authors. Australian Veterinary Journal © 2013 Australian Veterinary Association.)

Fusobacterium necrophorum, a Gram-negative, anaerobic bacterium, is an opportunistic animal and human pathogen that causes a variety of infections termed necrobacillosis. There are 2 subspecies of F. necrophorum (subsp. necrophorum and subsp. funduliforme) that differ morphologically and biochemically and in virulence. Leukotoxin, a secreted protein, is considered to be the major virulence factor. In camelids, F. necrophorum causes a variety of infections, generally involving the lips, tongue, pharynx, interdigital spaces, foot pad, larynx, mandible, or maxillary bones. The objective of the current study was to characterize the presumptive Fusobacterium isolates from a variety of necrotic infections in llama (Lama glama) and alpaca (Vicugna pacos) and determine whether the strains possess leukotoxin activities. A total of 7 isolates from alpaca and 2 isolates from llama were characterized. Based on growth characteristics in broth culture, and biochemical and polymerase chain reaction analyses, all 9 isolates belonged to subsp. necrophorum and possessed the putative hemagglutinin gene. Western blot analysis with antileukotoxin antibodies raised in rabbit showed the presence of leukotoxin protein in the culture supernatant of all isolates. Furthermore, flow cytometry of the culture supernatants demonstrated cytotoxicity to bovine and alpaca polymorphonuclear leukocytes (PMNs). The extent of cytotoxicity to either alpaca or bovine PMNs differed among camelid strains. The cytotoxicity of many of the camelid strains was higher (P < 0.05) toward alpaca PMNs compared to bovine PMNs. Fusobacterium necrophorum isolates from llama and alpaca are similar to bovine isolates, and leukotoxin may be a major virulence factor.

Members of the genus Actinobacillus comprise a diverse group of bacteria associated with mammals and birds including both pathogens and commensals. Here we describe the isolation of a previously undescribed Actinobacillus-like organism from seven epidemiologically unrelated infections of alpaca. The isolates are phenotypically and genotypically distinct from any previously described Actinobacillus species but 16S rRNA analysis unequivocally places the isolates as a novel lineage within the Actinobacillus sensu stricto. The clinical relevance of the organism requires further study however isolation in pure culture from organs of some cases suggests it may be associated with septicaemia in juvenile alpaca., (Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.)

This is the first report of detection of Candidatus Mycoplasma haemolamae in alpacas in England. The primary case occurred in a three year-old male alpaca in the south-east of England which presented with a history of progressive weight loss, lethargy, swelling of the scrotum and pale mucous membranes. Blood smear examination revealed a moderate, regenerative anaemia, with numerous small basophilic coccoid structures consistent with Candidatus M haemolamae. To confirm the presence of Candidatus M haemolamae, a portion of the 16S rDNA gene was amplified and analysed by denaturing gradient gel electrophoresis (DGGE). 16S rDNA gene sequencing showed a 99.8 per cent homology with Candidatus M haemolamae sequences deposited in GenBank. Subsequently, a cross-sectional study was carried out to investigate the presence of Candidatus M haemolamae infection in the alpaca herd from which the primary case was detected (n=131). Blood smear examinations and PCR with DGGE were used and compared with a species-specific PCR. The prevalence of infection when PCR positive results were combined was 29 per cent. A substantial agreement between the PCR/DGGE and the species-specific PCR was found (κ=0.86). A significant association was also found between age and infection (P=0.04) while no significant association was found with sex or origin.

In this work we report the genome of Corynebacterium pseudotuberculosis strain 267, isolated from a llama. This pathogen is of great veterinary and economic importance, as it is the cause of caseous lymphadenitis in several livestock species around the world and causes significant losses due to the high cost of treatment.

A 6-hour-old alpaca was presented for evaluation of respiratory difficulty. As part of routine surveillance, methicillin-resistant Staphylococcus aureus (MRSA) was identified from a nasal swab taken upon admission to the hospital. No signs of MRSA infection were noted. The MRSA strain recovered was a human epidemic clone that has been associated with horses. Methicillin-resistant S. aureus colonization can occur in camelids, and the potential animal and public health risks require consideration.

Forty-two isolates of Streptococcus pluranimalium were identified from cattle (n=38), sheep (n=2), an alpaca (n=1) and a pheasant (n=1) in the United Kingdom. The isolates were confirmed as S. pluranimalium by 16S rRNA sequence analysis but could not be differentiated reliably from Streptococcus acidominimus by phenotypic characterisation using commercial kits routinely used in veterinary laboratories. The alanyl-phenylalanyl-proline arylamidase reaction could be used to differentiate S. pluranimalium (positive) from Aerococcus urinae (negative)., (Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.)

A mature female alpaca was evaluated for weight loss and a 10-day history of anorexia, diarrhea, abdominal distension, and ventral edema. Ultrasonography revealed a hepatic mass, culture of which identified Corynebacterium pseudotuberculosis. This is the first reported case of an internal caseous lymphadenitis lesion resulting in clinical disease in a camelid.

In spite of great efforts for its control and eradication, tuberculosis remains one of the most important zoonosis worldwide. Its causative agents, the members of the Mycobacterium tuberculosis complex, have a wide host range that complicates the epidemiology of this disease. Among susceptible species to these pathogens, camelids from the New World (llama, alpaca and vicuña) and Old World (Bactrian camel and dromedary) are acquiring an increasing importance in several European countries because of its growing number and could act as reservoirs of the disease for livestock and humans in their natural habitat. In addition, tuberculosis caused by a number of M. tuberculosis complex members is a life-threatening disease in these animal species. Although tuberculosis has been known to affect camelids for a long time, ante-mortem diagnosis is still challenging because of the lack of standardized diagnostic techniques and the limited sensitivity and specificity of the most widely applied tests. However, in recent years, several techniques that can at least partially overcome these limitations have been developed. This paper reviews the results and advances achieved in tuberculosis diagnosis in camelids in the last decade as well as the progresses on ongoing investigations, with special attention to the remaining challenges that still have to be faced to assure the availability of reliable tools for the detection of tuberculosis-infected animals and herds., (© 2011 Blackwell Verlag GmbH.)

Background: Methanogens that populate the gastrointestinal tract of livestock ruminants contribute significantly to methane emissions from the agriculture industry. There is a great need to analyze archaeal microbiomes from a broad range of host species in order to establish causal relationships between the structure of methanogen communities and their potential for methane emission. In this report, we present an investigation of methanogenic archaeal populations in the foregut of alpacas., Results: We constructed individual 16S rRNA gene clone libraries from five sampled animals and recovered a total of 947 sequences which were assigned to 51 species-level OTUs. Individuals were found to each have between 21 and 27 OTUs, of which two to six OTUs were unique. As reported in other host species, Methanobrevibacter was the dominant genus in the alpaca, representing 88.3% of clones. However, the alpaca archaeal microbiome was different from other reported host species, as clones showing species-level identity to Methanobrevibacter millerae were the most abundant., Conclusion: From our analysis, we propose a model to describe the population structure of Methanobrevibacter-related methanogens in the alpaca and in previously reported host species, which may contribute in unraveling the complexity of symbiotic archaeal communities in herbivores.

The breeding of wild animals for commercial purposes is becoming more frequent nowadays. This situation has led to an increase in contact rates between wild and domestic animals, with subsequent reciprocal transmission of parasites. In this study, we characterized the gastrointestinal and blood parasites of a group of 15 semi-captive guanacos (Lama guanicoe). We characterized gastrointestinal parasites by analyzing fecal samples through the sedimentation-flotation technique and hemoparasites by using blood smears stained with Giemsa. We found several gastrointestinal parasites including Nematoda and protozoans. The most frequently found parasites were Nematodirus sp. and Eimeria sp. In contrast with previous studies, neither Cestoda nor Fasciola were found. The only hemoparasite detected was Mycoplasma haemolamae, a parasite already described in llamas and alpacas. We conclude that the most frequent gastrointestinal parasites of semi-captive guanacos were nematodes and protozoans. Also, the hemoparasite M. haemolamae seems to be prevalent among captive populations of South American camelids. Finally, captive guanacos share several parasites with the traditional livestock. Therefore, keeping captive or semi-captive guanacos without an adequate sanitary protocol might have adverse consequences to adjacent traditional cattle farming and/or for wild animals.

Two alpacas from a herd in southwest Switzerland died for unknown reasons. Necropsy revealed chronic weight loss and pale mucous membranes. Infection with hemotropic mycoplasmas was suspected and subsequently confirmed by molecular methods. In order to investigate the epidemiological situation in this herd, a real-time TaqMan(®) qPCR assay for the specific detection and quantification of hemoplasma infection in South American camelids was developed. This assay was based on the 16S rRNA gene and amplified 'Candidatus Mycoplasma haemolamae' DNA, but not DNA from other hemoplasmas or non-hemotropic mycoplasma species. The lower detection limit was one copy/PCR, and the amplification efficiency was 97.4%. In 11 out of 24 clinically healthy herd mates of the two infected alpacas, 'Candidatus M. haemolamae' infection was confirmed. No correlation was found between bacterial load and clinical signs or anemia. The assay described herein enables to detect and quantify 'Candidatus M. haemolamae' and may be used in future studies to investigate the prevalence, pathogenesis and treatment follow-up of hemoplasma infections in South American camelids., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Vancomycin-resistant enterococci are a major cause of nosocomial infections but are rarely found in humans in the community and have not been identified in food animals in the United States. We evaluated a total of 360 fecal specimens from humans and their animals being raised for exhibit at three county fairs in Michigan. Fecal samples from 158 humans, 55 swine, 50 cattle, 25 horses, 57 sheep, 14 goats, and 1 llama were obtained and plated onto Enterococcosel agar containing 16 μg/ml of vancomycin. Vancomycin-resistant Enterococcus faecium (VREF) was isolated from six pigs but not from humans or any animal other than pigs. All six VREF isolates had a MIC to vancomycin of ≥256 μg/ml and contained the vanA gene. Pulsed-field gel electrophoresis (PFGE) patterns of the six VREF isolates were ≥80% similar. Multilocus sequence typing (MLST) revealed sequence type 5 (ST5) (n = 2), ST6 (n = 3), and ST185 (n = 1), which are E. faecium sequence types belonging to clonal complex 5 (CC5). These findings show the dissemination of VREF strains among pigs in three Michigan counties. This is the first report of VRE found in food animals in the United States.

Haemotrophic mycoplasmas (also known as haemoplasmas), small bacterias which parasite the surface of erythrocytes, have been described in several species. Recently, molecular methods were developed for the diagnosis of haemoplasma infection. The presented study describes the first detection and the investigation of prevalence of "Candidatus Mycoplasma haemolamae" in South American Camelids in Switzerland. A random sample of the latter population was tested for haemoplasma infections using real-time PCR. The infection was detected in 18.6% of the animals and was found both in indigenous and in imported camelids. Of the tested herds 39,1% harboured at least one animal positive for haemoplasmas in PCR. There was no difference in prevalence between male and female animals and llamas and alpacas, respectively. Furthermore, the prevalence of infection was not significantly different in diseased animals compared to healthy camelids. From the latter observation and the fact that the high prevalence was accompanied by an undetectable incidence, we concluded that the pathogenicity of "Candidatus Mycoplasma haemolamae" may be low.

An outbreak of tuberculosis (TB) caused by Mycobacterium bovis in a llama herd is described. Over a 25-month period, a total of 70 llamas were selected for postmortem examination using four distinct criteria: clinical suspicion of disease (15 animals), positive tuberculin skin test result (three animals), antibody positive using a novel serological test (Rapid Test, 54 animals) and elective cull (five animals). Some animals qualified on more than one criterion. Gross lesions of TB were detected in 15 animals, with lung and lymph node lesions consistently observed. Samples were collected from 14 of 15 animals with visible lesions as well as those with no visible lesions, for histopathology and mycobacterial culture. All 14 llamas with visible lesions had caseonecrotic granulomatous lesions associated with acid-fast bacteria and variable mineralisation, and M bovis was isolated from 13. There were no histopathological lesions of TB in llamas with no grossly visible lesions, and M bovis was not isolated from any of these. The predictive value of suspicious gross lesions at postmortem examination was therefore high in the herd. Molecular typing results indicated that the outbreak was caused by a single strain likely to have originated from a local reservoir, probably cattle or wildlife. Antemortem indicators of infection assisted control of the outbreak, but no single test accurately identified all TB cases. Visible lesions were detected in nine of 15 llamas with clinical suspicion of disease, in two of three that had positive tuberculin skin test results and in 10 of 54 that were antibody positive; there was none (zero out of five) in llamas that were electively culled.

Mycoplasma haemolamae is a hemotropic mycoplasma that affects red blood cells of llamas (Lama glama) and alpacas (Lama pacos). It is variably associated with anemia, and most infections are subclinical. Development of a polymerase chain reaction assay has facilitated detection of this infection in llamas and alpacas in the United States and other countries. Whether the infection occurs in camelids in South America has previously been unknown. The current study documents a 15.8% infection rate among 76 Peruvian llamas, a 19.3% infection rate among Peruvian alpacas at one site, and a 9.26% infection rate in 108 Chilean alpacas from selected herds. All of the camelids tested appeared to be clinically healthy. No gender or species predilection was found. Only 1 positive camelid younger than 18 months was found. Infection is not associated with anemia, and the mean packed cell volume (PCV) in positive Peruvian camelids was slightly higher than the mean PCV in negative Peruvian camelids. In the Chilean alpacas, the positive alpacas had a slightly lower PCV than the negative alpacas, although the mean PCV was not in the anemic range in any of the groups.

Two bacterial 16S rRNA gene clone libraries were constructed from the forestomach of alpacas and sheep fed alfalfa. After the amplification using the universal 16S rRNA gene primers, equal quantities of PCR products from the same species were mixed and used to construct the two libraries. Sequence analysis showed that the 60 clones from alpacas were divided into 27 phylotypes with 25% clones affiliated with Eubacterium sp. F1. The 60 clones from sheep were divided into 21 phylotypes with 7 phylotypes affiliated with Prevotella ruminicola (40% clones). Clones closely related to Clostridium proteoclasticum, Eubacterium sp. F1, Clostridium cellobioparum, Mogibacterium neglectum, Eubacterium ventriosum, Clostridiaceae bacterium WN011, Clostridium coccoides, Clostridium orbiscindens, Eubacterium sp. F1, Cytophaga sp. Dex80-37, Treponema bryantii and Pelotomaculum sp. FP were only found in the forestomach of alpacas, and those to Anaerovorax odorimutans, Treponema zioleckii, Bifidobacterium indicum, Paludibacter propionicigenes, Paraprevotella clara, Eubacterium siraeum, Desulfotomaculum sp. CYP1, Clostridium bolteae, Clostridium termitidis and Clostridiaceae bacterium DJF&#95;LS40 only in the rumen of sheep. Quantitative real-time PCR revealed that the forestomach of alpacas had significantly lower density of bacteria, with bacterial 16S rRNA gene copies (6.89 [Log10 (copies per gram of wet weight)]), than that of sheep (7.71, P<0.01). The two clone libraries also appeared different in Shannon index (library from alpacas 3.30 and from sheep 3.04). Our results showed that there were apparent differences in the bacterial diversity and abundance in the forestomach between alpacas and sheep., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)

A 44-year-old woman who had recently been on immunosuppressive therapy presented with malaise, cough, fever, weight loss, lymphadenopathy, severe hypercalcaemia and a paratracheal mass on imaging. The initial impression was of disseminated malignancy, and lymphoma was suspected. A mediastinal biopsy showed a mycobacterial spindle cell pseudotumour containing acid and alcohol fast bacilli (AAFB). Sputum microscopy demonstrated AAFBs, confirmed as Mycobacterium tuberculosis complex by PCR. Prolonged culture grew Mycobacterium microti, an organism often associated with disease in small rodents and llamas. M microti isolates from postmortem samples of an alpaca at a nearby farm were genetically indistinguishable. Although the patient had not visited the farm, concurrent illness in her adopted stray cat suggested a possible zoonotic connection. The patient responded to antituberculous therapy, and rehydration and pamidronate for hypercalcaemia. We believe the hypercalcaemia was caused by a similar mechanism to raised calcium levels sometimes seen in tuberculosis.

We report three cases of tuberculosis in alpacas from Spain caused by Mycobacterium bovis. The animals revealed two different lesional patterns. Mycobacterial culture and PCR assay yielded positive results for M. bovis. Molecular typing of the isolates identified spoligotype SB0295 and identical variable-number tandem repeat (VNTR) allele sizes.

A 7-year-old male llama was examined for a 3-month history of weight loss, and unilateral keratouveitis. Clinical examination revealed nonulcerative corneal stromal abscessation, corneal vascularization, corneal edema, miosis, posterior synechia, cataract, and fibrin in the anterior chamber of the right eye. The left eye was normal. Histopathology of the right eye following enucleation revealed pyogranulomatous keratouveitis with intralesional fungal spherules consistent with Coccidioides spp. PCR amplification with DNA sequencing confirmed Coccidioides posadasii infection. To the authors' knowledge, this is the first reported case of ocular coccidioidomycosis in a llama.