DNA barcoding for identification of cryptic species in the field and existing museum collections: a case study of Aethomys and Micaelamys (Rodentia: Muridae)

DNA barcoding has been proposed as a method for species identification. However, this method has been criticised for its over-reliance on a single mitochondrial gene. In this study, four mitochondrial gene regions and one nuclear gene region were used to investigate their different abilities to identify tissue associated with museum specimens of Aethomys chrysophilus, Aethomys ineptus and Micaelamys namaquensis. Aethomys chrysophilus and the more recently elevated A. ineptus are indistinguishable on morphological grounds; however, their ranges are largely parapatric with only one syntopic locality currently known. All of the mitochondrial gene regions were able to separate M. namaquensis from A. chrysophilus and A. ineptus, but they varied in their abilities to resolve differences between A. chrysophilus and A. ineptus. The sequence results identified a specimen from KwaZulu-Natal that was misclassified and should have been identified as A. ineptus. Seven specimens that had not been reclassified following the elevation of A. ineptus to species level were identified as A. ineptus. Individuals of A. chrysophilus from Malawi could not be classified as either A. chrysophilus or A. ineptus, and may be a hybrid or a new, distinct species. This study indicates that DNA barcoding may be used to separate M. namaquensis from A. chrysophilus and A. ineptus, and although it was not able to separate A. chrysophilus and A. ineptus, it did indicate specimens from Malawi may be a new cryptic species.     

Taxonomic research on deep‐sea macrofauna in the South China Sea using the Chinese deep‐sea submersible Jiaolong

This paper reviews the taxonomic and biodiversity studies of deep‐sea invertebrates in the South China Sea based on the samples collected by the Chinese manned deep‐sea submersible Jiaolong. To date, 6 new species have been described, including the sponges Lophophysema eversa, Saccocalyx microhexactin and Semperella jiaolongae as well as the crustaceans Uroptychus jiaolongae, Uroptychus spinulosus and Globospongicola jiaolongi; some newly recorded species from the South China Sea have also been reported. The Bathymodiolus platifrons–Shinkaia crosnieri deep‐sea cold seep community has been reported by Li (2015), as has the mitochondrial genome of the glass sponge L. eversa by Zhang et al. (2016). The population structures of two dominant species, the shrimp Shinkaia crosnieri and the mussel Bathymodiolus platifrons, from the cold seep Bathymodiolus platifrons–Shinkaia crosnieri community in the South China Sea and the hydrothermal vents in the Okinawa Trough, were compared using molecular analysis. The systematic position of the shrimp genus Globospongicola was discussed based on 16S rRNA gene sequences.     

Distribution of Rhodococcus equi in animals,birds and from the environment

Extract

Madam:— As well as causing sporadic infections in animals, Rhodococcus (Corynebacterium) equi has been isolated from the gastro-intestinal tract of healthy grazing animals and from the environment. ^{(1 )(5)(6)(8)(11)} R. equi has been isolated in this laboratory from cattle and pigs in association with tuberculosis- like lesions and from lungs and abscesses from horses and deer.     

Comparative Susceptibility of Channel Catfish,Blue Catfish,and their Hybrid Cross to Experimental Challenge with Bolbophorus damnificus (Digenea: Bolbophoridae) Cercariae

Abstract

The digenetic trematode Bolbophorus damnificus has been implicated in significant losses in catfish aquaculture since the late 1990s. The complex life cycle sequentially involves the American white pelican Pelecanus erythrorhynchos, the marsh rams horn snail Planorbella trivolvis, and Channel Catfish Ictalurus punctatus. Research supports anecdotal reports from the industry, suggesting that the hybrid of Channel Catfish×Blue Catfish I. furcatus is less susceptible to disease agents that have been historically prohibitive to Channel Catfish production, namely the gram-negative bacteria Edwardsiella ictaluri and Flavobacterium columnare, as well as the myxozoan parasite Henneguya ictaluri. This current research compared the susceptibility of Channel Catfish, Blue Catfish, and their hybrid cross to an experimental challenge by B. damnificus. Fish were exposed to 0, 100, 200, and 400 B. damnificus cercariae per fish, and the numbers of metacercariae per fish were determined 14 d postchallenge. Metacercariae were recovered from all challenged fish. There were no significant differences among fish groups challenged with the same dose, suggesting Channel and Blue Catfish and their hybrid are comparably susceptible to B. damnificus infection. As such, it is recommended that producers raising hybrid catfish remain diligent in controlling populations of the snail intermediate host to prevent production losses attributed to B. damnificus, especially when loafing pelicans have been observed at the aquaculture operation.

Received October 22, 2013; accepted January 5, 2014     

Occurrence of avian Schistosomatidae (Trematoda) in South African birds as determined by a faecal survey

The abundance, prevalence and distribution of avian Schistosomatidae in South African birds has been estimated by means of a survey for parasite eggs in faecal samples. Eight types of eggs were recovered, mostly from members of the Anatidae and Laridae and these have been assigned to the following schistosome genera: Austrobilharzia (1), Gigantobilharzia (1), Trichobilharzia (5) and Ornithobilharzia (1).     

Presence of Ureaplasma diversum in the Australian cattle population

In cattle, Ureaplasma diversum has been associated with decreased fertility, granular vulvovaginitis, endometritis, salpingitis and spontaneous abortion in cows and seminal vesiculitis, balanoposthitis and changes in bull sperm. The presence of U. diversum within the Australian cattle population has not been established. One of the aims of this study was to determine if U. diversum was present in Australian cattle, using culture and polymerase chain reaction (PCR), both of which are considered to be gold standards for bacterial identification. Of 100 samples collected from 66 male and 34 female cattle, 15 were positive for U. diversum. Therefore, Australia can no longer be considered free of U. diversum. Further studies should be conducted to ascertain the effects of U. diversum within Australian cattle herds and, if warranted, to investigate prevention, treatment and eradication protocols.     

Faecal shedding and serological cross‐sectional study of Lawsonia intracellularis in horses in the state of Minas Gerais,Brazil

Reason for performing the study: Proliferative enteropathy, caused by the intracellular bacterium Lawsonia intracellularis, has been described in horses in Australia, the USA, Canada and European countries but has not been reported in Latin America. The prevalence of the disease in horses worldwide is unknown. Objective: To evaluate the presence of subclinical L. intracellularis infection in horses in the state of Minas Gerais, Brazil. Methods: A longitudinal study using serology and PCR for detecting antibodies (IgG) and shedding of L. intracellularis in faecal samples, respectively, was conducted using a total of 223 horses from 14 different horse farms in Minas Gerais, and from the Veterinary School of UFMG equine herds in Minas Gerais. The immunoperoxidase technique in glass slides was used as the serological test. Results: Twenty‐one horse sera had immunoglobulin G titres of 1:60 and were considered positive. The PCR technique in faeces for L. intracellularis DNA identified 7 horses as faecal shedders. Horses shedding the organism appeared healthy, indicating that subclinical infection of L. intracellularis occurred in the horses. Conclusion: Seropositivity and detection of faecal shedding of L. intracellularis indicates the presence of the agent in the equine population in Minas Gerais. Potential relevance: Results of this study should alert clinicians in countries where proliferative enteropthy in horses has not been reported to consider this disease as a possible cause of enteric disease.     

Ingestion of host immunoglobulin by three non-blood-feeding nematode parasites of ruminants

Host immunoglobulin was detected in homogenates of adult worms or fourth stage larvae of Haemonchus contortus, Ostertagia ostertagi, O circumcincta and Dictyocaulus viviparus, but not in preparasitic third stage larvae of H contortus. All the worms had been washed in detergent before being homogenised to remove any immunoglobulin adhering to the cuticle. This procedure, the concentrations of immunoglobulin detected and observations on sections of the worms stained with fluorescent antisheep immunoglobulin, all suggested that the immunoglobulin had been ingested, even by the non-blood-feeding species. This finding suggests that the Ostertagia species and D viviparus may be susceptible to vaccination by the gut antigen approach, a method with considerable promise for blood-feeding parasites like H contortus and Boophilus microplus.     

Prevalence of Yeasts and Mycelial Fungi in Bovine Parasitic Otitis in the State of Minas Gerais,Brazil

Infestations by rhabditiform nematodes and acarids of the genus Raillietia are considered the primary causes of external otitis in cattle in tropical regions. Recently, yeasts of the genus Malassezia have been associated with a relatively high percentage of otitis cases, but the occurrence of other yeasts and mycelial fungi has not yet been reported in the literature. This work studied the presence of fungi in the ear canal of 45 cattle with external parasitic otitis. The results were positive for yeasts of the genus Malassezia in 31 (68.9 %) of the 45 cultures in Mycosel medium supplemented with olive oil. The 45 cultures in Sabouraud dextrose medium revealed the growth of seven (15.5 %) yeasts of the genus Candida, five (11.1 %) Rhodotorulamucilaginosa, two (4.4 %) fungi of the genus Aspergillus and eight ‘Micelia sterilia’. Future studies may confirm and elucidate the importance of these agents in the aetiology of bovine otitis.     

Antimicrobial resistance in bacteria from horses: Epidemiology of antimicrobial resistance

Antimicrobial resistance poses a significant threat to the continued successful use of antimicrobial agents for the treatment of bacterial infections. While the epidemiology of antimicrobial resistance in bacteria from man has been studied extensively, less work has been undertaken in companion animals, particularly horses. Methicillin‐resistant Staphylococcus aureus has been identified as a cause of infections, with a low prevalence of nasal carriage by horses in the community but higher for hospitalised horses. Molecular characterisation has shown methicillin‐resistant Staphylococcus aureus strains either to be predominantly of types associated with horses or of sequence type ST398. Antimicrobial‐resistant Escherichia coli (including multidrug‐resistant and extended spectrum β‐lactamase‐producing isolates) have caused infections and been documented in faecal carriage by horses, with many significant resistance mechanisms identified. More sporadic reports and molecular characterisation exist for resistance in other bacteria such as enterococci, Salmonella, Acinetobacter and Pseudomonas species. Limited work has been undertaken evaluating risk factors and much of the epidemiology of antimicrobial resistance in bacteria from horses remains to be determined.     

Caprine sex affects skeletal muscle profile and MRFs expression during postnatal development

The important roles of myogenic regulatory factors (MRFs) have been well addressed in the process of mammalian skeletal myogenesis, while limited research has been performed in small ruminants. Furthermore, the effects of gender on the development of skeletal muscle and MRFs expression remain unknown. In this study, we identified the caprine Myf5, Myf6, MyoD and myogenin genes and quantified their expressions at six different postnatal time points by real‐time RT‐PCR. The sex has a marked effect on the expression differences of Myf5, MyoD and myogenin in the five investigated skeletal muscles, while minor influence on the expression difference of Myf6 except for Semitendinosus and Quadriceps femoris tissues (P < 0.001). The histological sections of muscles revealed a constant increase of muscle fiber diameter with aging but non‐significant differences between genders. We provide novel evidence for MRFs expression in age‐ and gender‐dependent manners, which will contribute to prioritizing these genes as potential candidate genes for trait‐associated study and provide a foundation for understanding the molecular control of skeletal muscle growth in goat species.     

Global Distribution and Prevalence of Arcobacter in Food and Water

The emerging foodborne and waterborne pathogen, Arcobacter, has been linked to various gastrointestinal diseases. Currently, 19 species are established or proposed; consequently, there has been an increase in the number of publications regarding Arcobacter since it was first introduced in 1991. To better understand the potential public health risks posed by Arcobacter, this review summarizes the current knowledge concerning the global distribution and the prevalence of Arcobacter in food and water. Arcobacter spp. were identified in food animals, food‐processing environments and a variety of foods, including vegetables, poultry, beef, dairy products, seafood, pork, lamb and rabbit. A wide range of waterbodies has been reported to be contaminated with Arcobacter spp., such as wastewater, seawater, lake and river water, drinking water, groundwater and recreational water. In addition, Arcobacter has also been isolated from pets, domestic birds, wildlife, zoo and farm animals. It is expected that advancements in molecular techniques will facilitate better detection worldwide and aid in understanding the pathogenicity of Arcobacter. However, more extensive and rigorous surveillance systems are needed to better understand the occurrence of Arcobacter in food and water in various regions of the world, as well as uncover other potential public health risks, that is antibiotic resistance and disinfection efficiency, to reduce the possibility of foodborne and waterborne infections.     

Local-scale diversity of Yersinia pestis: A case study from Ambohitromby,Ankazobe District,Madagascar

Plague is a re-emerging zoonotic disease and a major public health concern in several portions of the world, especially in Madagascar. We report on the presence of different subtypes of Yersinia pestis co-occurring in the same locality. After confirmation of a human plague case in Ambohitromby Commune (Ankazobe District) via isolation of Y. pestis, we undertook small mammal trapping to identify the circulation of Y. pestis amongst rodents in this locality; blood samples were collected from rodents for seroprevalence analysis. Of the 60 individuals of Rattus rattus captured, one yielded an isolate of Y. pestis, 13 others were positive for F1 antigen of Y. pestis using a rapid diagnostic test, and 4 were PCR positive targeting the caf1 and pla genes; 28/60 (46.7%) of the captured R. rattus were seropositive for Y. pestis. Whole-genome SNP analyses revealed that the two isolates obtained from the human case, and the R. rattus belonged to two different subtypes of Y. pestis (s05 and s13, respectively) that were circulating concurrently in Ambohitromby in 2016. Three Y. pestis subtypes (s03, s05 and s13) have now been isolated from Ambohitromby. Subtype s05 had been persisting there for >10 years but one or both of the other subtypes may have been introduced from the Central Highlands region as they were not observed in previous years (s13) or only observed once previously (s03). High seroprevalence against Y. pestis in R. rattus suggests that a portion of the local murine population may have acquired resistance to Y. pestis. Future research should focus on genomically characterizing Y. pestis strains circulating in Ankazobe District and other plague-endemic regions of Madagascar to better understand the overall phylogeography of Y. pestis.     

Microsporidiosis: Enterocytozoon bieneusi in domesticated and wild animals

Microsporidia are a ubiquitous group of obligate intracellular parasites that infect all major animal groups. Enterocytozoon bieneusi is the most commonly identified Microsporidia in humans and has also been reported worldwide in animals with importance in veterinary medicine (e.g. cats, dogs, horses, cattle and pigs). The identification of E. bieneusi in animals has raised the question of the importance of animal reservoirs in the epidemiology of this pathogen, and the implications of the infection with this pathogen in infected animals. Considerable genetic diversity within E. bieneusi has been found with over 90 genotypes identified based on the ITS nucleotide sequence of E. bieneusi spores recovered from the feces of infected humans and animals. Both host-adapted E. bieneusi genotypes with narrow host ranges and potentially zoonotic genotypes with wide host specificity have been identified. The information presented in this review should be useful in understanding the taxonomy, epidemiology, zoonotic potential, and importance in public health of E. bieneusi.     

Prognostic value of tumour‐associated macrophages in canine mammary tumours

Tumour‐associated macrophages (TAMs) have already been associated in human breast cancer to a poor prognosis. As a part of a tumoural microenvironment, TAMs have an important contribution influencing neoplastic progression. Hitherto, in canine mammary tumours (CMT) the prognostic value of TAMs has not been reported. In this study, MAC387 immunohistochemical expression was evaluated in 59 CMTs (20 benign and 39 malignant). The TAM value was significantly higher in malignant than benign CMT (P = 0.011). In malignant CMT, TAMs were associated with skin ulceration (P = 0.022), histological type (P = 0.044), nuclear grade (P = 0.031) and tubular differentiation (P = 0.042). The survival analysis revealed a significant association between tumours with higher levels of TAMs and the decrease in overall survival (P = 0.030). TAMs have proven to have a prognostic value. These findings suggest the future possibility of using TAMs as a novel therapeutic target in CMT.     

The distribution of Nematodirus spp. occurring in sheep in New Zealand

Extract

Most outbreaks of parasitic gastro-enteritis in sheep in New Zealand have been ascribed to the presence of nematodes of the family Trichostrongylidae, and in particular the genera, Haemonchus, Ostertagia, and Trichostrongylus. However, during the period 1956–58, a number of outbreaks of disease occurred in lambs and the condition was associated with heavy Neniatodirus infestation. Although Neinatodirus spp. are commonly foundin sheep in this country, only moderate numbers of worms are usually present in any individual and the worm has been regarded as non-pathogenic. Similar views were held in the British Isles until about 1951, when an acute disease condition of lambs, associated with heavy Neniatodirus burdens, caused serious losses.     

Hyperolius argus (Anura) in Natal: taxonomy,biogeography and conservation

A high degree of variation in the colour pattern of Hyperolius argus Peters, including sexual dichromatism and a marked cline, has led to much taxonomic confusion. This East African species extends down the Natal coastal lowlands as far south as Durban. It has been assigned to H. punctlculatus (Pfeffer) in Natal. In a discussion of Mozambique and Natal material, this assignation is shown to be incorrect. The probable holotype of Cope's cinctiventrls is considered to be a synonym of argus. There is some evidence that populations of argus intergrade with semidiscus Hewitt in Natal, and the occurrence of semidiscus on the periphery of the argus range is discussed in relation to the ‘central-marginal’ model of biogeographical patterning. It has not been possible to determine the relative vulnerability to habitat disturbance of argus compared to any other amphibian species, but the conservation prospects of this species at the southern end of its range currently seem extremely poor