Treatment of rats with heat killed cells (HKC) of Cryptococcus neoformans var. grubii induces cellular activation in spleen and lymphatic nodes

Our previous studies showed that the subcutaneous pretreatment of rats with heat killed cells of Cryptococcus neoformans (HKC) emulsified in complete Freund adjuvant (CFA) promotes protection against an intraperitoneal challenge with viable C. neoformans. In this model, an appropriate activation of adherent peritoneal cells after antigenic treatment is very important for the control of the infection. Here, we investigated the immune response developed in spleen and lymphatic nodes as a result of treatment with HKC-CFA, which might also contribute in the protective phenomenon of this treatment against cryptococcal infection. The results show that, compared with adjuvant alone, rats which received treatment with HKC-CFA presented a greater activation of adherent splenic cells, with up-regulation of major histocompatibility complex class II (MHC II) and CD86 expression and secretion of anticryptococcal metabolites. Furthermore, this treatment also induced an increase in the blastogenic response and the secretion of Th1 and Th2 cytokines by spleen cells in comparison with cells from CFA-phosphate-buffered saline (PBS) treated rats. On the other hand, lymph node cells from animals treated with HKC-CFA presented a rise in the expression of MHCII but not of CD86 with respect to control cells from rats treated with CFA-PBS. These cells also showed a high proliferative response and secretion of Th1-related cytokines, interleukin (IL)-12 and tumor necrosis factor (TNF). These results show that treatment of rats with HKC-CFA is able to induce an early immune response in secondary lymphoid organs, which may contribute to the protective effect induced by this treatment.     

Immobilization of free-ranging maned wolf (Chrysocyon brachyurus) with tiletamine and zolazepam in central Brazil.

A tiletamine hydrochloride-zolazepam hydrochloride combination was used successfully to immobilize 27 free-ranging maned wolves (Chrysocyon brachyurus) at a mean dose of 2.77+/-0.56 (mean+/-SD) mg/kg. The induction time ranged from 3-15 min. Animals remained immobilized for periods of 48.56 +/-12.65 min. Compulsive licking, excessive salivation, muscle twitching, muscle tremors, tachypnea, and bradycardia were observed associated with the induction of the anesthesia in 13 of 27 maned wolves. Muscle twitching, pedal withdrawal reflex, muscle tremors, and ataxia were observed during recovery in three (11%) maned wolves. There were no significant differences in heart rates (P = 0.44), respiratory rates (P = 0.82), and rectal temperatures (P = 0.54) recorded at 5, 15, and 25 min after induction at these dosages. The tiletamine hydrochloride-zolazepam hydrochloride combination was shown to be an effective and safe immobilizing agent for free-ranging maned wolves.     

Cryptococcus neoformans granuloma in the lung and spinal cord of a free-ranging cheetah (Acinonyx jubatus). A clinical report and literature review

A 6-year-old, male, wild-born, free-ranging cheetah (Acinonyx jubatus) was evaluated for acute onset of progressive lameness in the right hind limb. Survey radiographs were unrewarding and myelography indicated an intramedullary compressive mass at the L3-L4 region. A fine needle aspirate of the lesion indicated the presence of Cryptococcus organisms. Necropsy confirmed the presence of granulomas (cryptococcoma) in the lung and the spinal cord (meningomyelitis) caused by Cryptococcus neoformans var. gattii. Cryptococcus neoformans is a yeast-like organism that is a potential pathogen to many species. Initial infection is thought to be of respiratory origin and then it commonly disseminates systemically from the nasal cavity or lungs to the skin, eyes and central nervous system in particular. The cheetah tested negative for both feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV), as have all the previously reported cheetah cases. C. neoformans is a non-contagious, opportunistic organism and is the most common systemic mycoses in domestic cats and the cheetah.     

Isolation of culturable microorganisms from free-ranging bottlenose dolphins (Tursiops truncatus) from the southeastern United States

Reports of diseases in marine mammals are increasing worldwide, however our understanding of the microorganisms associated with marine mammals is still limited. In this study, we cultured bacteria and fungi isolated from the upper respiratory tract (blowhole), gastric fluid and anus of 180 wild bottlenose dolphins (Tursiops truncatus) from two estuarine locations along the southeastern Atlantic Coast of the United States. A total of 339 and 491 isolates from Charleston, SC (CHS) and Indian River Lagoon, FL (IRL) dolphins, respectively, were cultured from gastric (70 CHS/82 IRL), fecal (141 CHS/184 IRL), and blowhole (128 CHS/225 IRL) swabs on selective media used for routine clinical microorganisms of human concern. The most frequently cultured Gram-negative bacteria from all sample and study types were Plesiomonas shigelloides, Aeromonas hydrophila, Escherichia coli, and Pseudomonas fluorescens. Among the Gram-positive bacteria, Clostridium perfringens, Bacillus sp., and Staphylococcus Coag. Neg were the predominant organisms. For fungi, the most abundant species were Candida glabrata, budding yeasts, and Candida tropicalis. Of concern, the MRSA strain of Staphylococcus aureus was detected in the blowhole and gastric swabs from CHS dolphins. In general, a greater prevalence of bacteria and fungi (four-fold increase) were cultured from IRL than CHS animals. Together, these culture-dependent studies, coupled to on-going culture-independent approaches, should help establish a baseline of microorganisms associated with bottlenose dolphins and aid in the identification of organisms responsible for infectious diseases(s) in these animals.     

Isolation and genotyping of Campylobacter species from kiwi (Apteryx spp.) in captivity: implications for transmission to and from humans

Aims: To investigate the presence of Campylobacter spp. in captive kiwi (Apteryx spp.) and compare their genotypic profiles with those of human and animal origin, in order to assess their potential for zoonotic or zooanthroponotic transmission.

Methods: Conventional selective enrichment and filter-based isolation methods were applied to isolate Campylobacter spp. from fresh faecal samples from 12 North Island brown kiwi (Apteryx mantelli) and one great spotted kiwi (A. haastii), housed in one of five different areas in a kiwi sanctuary in Christchurch, New Zealand. Isolates were identified using multiplex PCR and 16S rRNA gene sequencing. High-resolution rapid genotyping using multiplex ligation-dependant probe amplification-based binary typing (MBiT) was applied and profiles compared with similar results from 2,165 Campylobacter spp. isolates contained in a database derived from human clinical, veterinary and environmental samples.

Results: One isolate of C. jejuni, and one belonging to the C. lari phylogenetic group were recovered from faeces from two kiwi. High-resolution rapid genotyping by MBiT demonstrated these to be indistinguishable from isolates obtained previously from human cases of diarrhoea, and others from chicken, cattle, sheep and water.

Conclusions: These data provide evidence for potential zoonotic or zooanthroponotic transmission of Campylobacter spp. in kiwi with implications for management of birds kept in captivity. We believe this is the first formal report of C. jejuni and a C. lari-like organism in kiwi.

Abbreviations: MBiT: Multiplex ligation-dependant probe amplification-based binary typing     

Exposure of free-ranging maned wolves (Chrysocyon brachyurus) to infectious and parasitic disease agents in the No?l Kempff Mercado National Park, Bolivia.

Maned wolves (Chrysocyon brachyurus) are neotropic mammals, listed as a CITES Appendix II species, with a distribution south of the Amazon forest from Bolivia, through northern Argentina and Paraguay and into eastern Brazil and northern Uruguay. Primary threats to the survival of free-ranging maned wolves include habitat loss, road kills, and shooting by farmers. An additional threat to the conservation of maned wolves is the risk of morbidity and mortality due to infectious and parasitic diseases. Captive maned wolves are susceptible to, and die from, common infectious diseases of domestic dogs (Canis familiaris) including canine distemper virus (CDV), canine parvovirus (CPV), rabies virus, and canine adenovirus (CAV). Results from this study show that free-ranging maned wolves in a remote area of Bolivia have been exposed to multiple infectious and parasitic agents of domestic carnivores, including CAV, CDV, CPV, canine coronavirus, rabies virus, Leptospira interrogans spp., Toxoplasma gondii, and Dirofilaria immitis, and may be at increased risk for disease due to these agents.     

Isolation and characterisation of local strains of Chlamydophila abortus (Chlamydia psittaci serotype 1) from Tunisia

Chlamydiosis is one of the major diseases that can lead to abortion in ewes. Since 1997, in 5 regions of Tunisia, Chlamydia-related abortions have been reported in 15 sheep and goat flocks. One hundred and sixty-six sera and 50 vaginal swab samples were collected from adult ewes. Chlamydial antigens were detected in 29 (58%) of the vaginal swabs using Enzyme Linked Immunosorbent Assay (ELISA) while 9 (18%) were positive by cell culture. Five strains were recovered from 4 different sheep flocks. Monoclonal antibody profiles and restriction fragment length polymorphism (RFLP) analysis of the 16S-23S rRNA spacer region showed that these isolates were C. abortus. Using amplified fragment length polymorphism (AFLP), these Tunisian strains were shown to exhibit the same pattern as strains isolated in France.     

兔须癣毛癣菌新变种的分离与鉴定

从暴发皮肤病的某家兔场患病兔中分离到1株病原真菌,直接镜检、分离培养及致病性试验后,得到一种致病真菌。通过培养特性、显微观察及分子鉴定,显示该菌培养特性与已报道须癣毛癣菌均不相同,所有培养基上均未见螺旋菌丝,ITS区进化树亦标明其与其他种(变种)均不在同一分支中,据此确定为须癣毛癣菌的新变种(Trichophyton mentagrophytes var.sp.)。     

Isolation,molecular identification,and phylogenetic evaluation of Cryptococcus neoformans isolated from pigeon lofts,Psittaciformes, and Passeriformes in Ahvaz,Iran

Cryptococcus neoformans, the main pathogen in immunocompromised patients, is a ubiquitous free-living fungus that can be isolated from avian excreta, soils, and plant material. This study was carried out to determine the infection rate of pigeon lofts, Passeriformes, and Psittaciformes in Ahvaz, the capital of Khuzestan province in Iran and to determine varieties of Cryptococcus neoformans (C. neoformans). The 80 samples were collected from pigeon lofts. Also, 163 feces of captive birds (Passeriformes and Psittaciformes) which kept in Ahvaz pet shops, and the 70 cloacal swabs of pet birds (Passeriformes and psittaciformes) referring to the department of avian medicine (the faculty of veterinary medicine of Shahid Chamran University of Ahvaz) were analyzed. The samples were directly inoculated on niger seed agar (NSA) and also enriched in brain heart infusion broth and then inoculated on NSA. Dark brown colonies suspected to C. neoformans subcultured on saborouds dextrose agar and pure cultures subjected to molecular (polymerase chain reaction (PCR)) diagnosis. For detection of C. neoformans, primer sets that targeting the CNLAC1 gene were selected and nested PCR was conducted. For identification of C. neoformans varieties, a primer set targeting the STR1 gene was selected. For more accurate confirmation, the purified PCR products of some isolates were also sequenced, and based on the gene sequences, all of the isolates belonged to C. neoformans variety grubii (var. grubii)(serotype A). Totally 16 out of 80 pigeon samples (20%) were contaminated with C. neoformans. The results in pigeons disclosed a 98.64% identity when compared with other strains of C. neoformans (CN1525, T4, and T1) which were previously deposited in GenBank from Italy and Thailand. Also, 21 out of 233 samples from Psittaciformes (9.01%) were contaminated with C. neoformans. The results in Psittaciformes disclosed a 99.7% identity when compared with other strains of C. neoformans (TIMM1313, IFM5882, CN1525, etc.) which were previously deposited in GenBank from Japan and Italy, etc. In the present study, the samples belonging to the passerine order were free of C. neoformans infection. According to the results, C. neoformans is prevalent in pigeon flocks and pet birds including Psittaciformes in the Ahvaz area, and should be considered by pigeon and captive bird breeders, veterinarians, and public health organizations in Ahvaz. The cryptococcus species isolated from captive birds and pigeons could be potential pathogens in humans.     

Osteology and radiology of the Maned Wolf (Chrysocyon brachyurus) pelvic limb

This study describes the osteology and radiology of the pelvic limb in maned wolves. Ten (five live and five dead) maned wolves (Chrysocyon brachyurus), five males and five females, aged from 2 to 7 years old were used. Digital radiographs were taken and recorded for both pelvic limbs in all animals. Osteology was correlated with the radiographic images. The pelvis had a rectangular shape, and the obturator foramen (foramen obturatum) was oval. The femoral neck (collum femoris) was short and thick. The greater trochanter (trochanter major) extended proximally to near the dorsum of the femoral head (caput ossis femoris). The lateral femoral condyle (condylus lateralis) was larger than the medial condyle (condylus medialis), and the intercondylar fossa (fossa intercondylaris) had a slightly oblique orientation. The proximal tibia displayed medial and lateral condyles with the medial larger. The femur was slightly shorter than the tibia. Seven tarsal bones (ossa tarsi) were present, four long metatarsal bones (ossa metatarsalia II ‐ V) and a short first metatarsal bone (os metatarsal I).     

药用植物柽柳内生真菌分离及其抑菌活性初步研究

为探明药用植物柽柳内生真菌资源的多样性及其抑菌活性特征,通过组织块法对内生真菌进行了分离,并选择小麦全蚀病菌、枸杞黑果病菌、番茄灰霉病菌、黄瓜枯萎病菌、黄瓜立枯病菌5种植物病原真菌和枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌4种细菌作为指示菌,采用对峙法和改进的菌块法测定其抑菌活性。结果显示,从柽柳不同组织器官中分离出18株内生真菌,茎部最多,根部次之,叶部最少;经形态学初步分类鉴定归于2个目,3个科,7个属;有11株菌对2种或多种植物病原真菌有不同程度的抑制作用,占菌株总数的61.1%,有8株内生真菌只对金黄色葡萄球菌具有明显抑制作用,占分离菌株总数的44.4%;活性内生真菌菌株TCEFL001分别对枸杞黑果病菌、小麦全蚀病菌以及金黄色葡萄球菌具有明显抑制作用,属于组丝核菌属。药用植物柽柳具有较为丰富的内生真菌资源,其开发和利用值得进一步探究。     

Isolation of Sarcocystis falcatula from the South American opossum (Didelphis albiventris) from Argentina.

Sarcocystis sporocysts from the intestines of four opossums (Didelphis albiventris) from Argentina were identified as Sarcocystis falcatula based on schizogonic stages and pathogenicity to budgerigars (Melopsittacus undulatus). Seven budgerigars fed sporocysts from the opossum feces died of acute sarcocystosis 8, 9, 11, 12, and 14 days after inoculation. Schizonts and merozoites found in the lungs and other organs of the budgerigars were identified as S. falcatula based on structure and immunoreactivity with S. falcatula-specific antibody. Sarcocystis falcatula was also isolated in bovine monocyte cell cultures inoculated with lung tissue from a budgerigar that died nine days after ingesting sporocysts. Two budgerigars inoculated subcutaneously with 1,000,000 culture-derived S. falcatula died 11 and 12 days post-inoculation. This is the first report of S. falcatula infection in South America.     

Viral diseases of the Ostrich (Struthio camelus var. domesticus)

The ostrich is an important animal in many livestock industries. A significant threat to this industry is losses from diseases. Newcastle disease is a notifiable, highly contagious viral infection of ostriches. Avian influenza may be transmitted from waterfowl, shorebirds and gulls to ostriches. Borna disease virus is a viral neurotropic infection spread mainly by rodents and felines. Crimean‐Congo hemorrhagic fever is a viral disease transmitted by Hyalomma ticks to humans. Avipoxvirus afflicts ostrich chicks and is transmitted by mosquitoes or by direct contact with a pox lesion. Maintenance of a healthy and profitable enterprise requires the implementation, with assistance from the local veterinary authority, of comprehensive, practical and effective methods of health management and preventative medicine.     

Collection and short-term preservation of semen from free-ranging eastern grey kangaroos (Macropus giganteus: Macropodidae)

Objectives To evaluate electro-ejaculation of free-range eastern grey kangaroos in the field and assess the efficacy of four diluents to preserve sperm motility over a 48-h period at 5°C.
Procedure and design Under gaseous anaesthesia, 25 free-range kangaroos were electro-ejaculated and characteristics of the ejaculate noted. Spermatozoa obtained from eight ejaculates were diluted in phosphate buffered saline containing various combinations of egg yolk and glucose and refrigerated at 5°C for 48 h.
Results Spermatozoa were recovered from 24 of 28 ejaculates. Mean (± SEM) semen volume (mL) and pH were 25.0 ± 1.9 and 7.1 ± 0.1 respectively. The forward motility (%), rate of movement of sperm (0 to 5) and sperm concentration (x 10⁶/mL) were 77.4 ± 1.5, 3.8 ± 0.9 and 31.2 ± 7.3 respectively. There was no significant difference between the four diluents in their ability to maintain forward motility of spermatozoa over 48 h. However, rate of movement over the same period was significantly (P < 0.01) improved when sperm were diluted in phosphate buffered saline containing 10% egg yolk.
Conclusions Electro-ejaculation is a safe and reliable method for collecting semen from free-ranging eastern grey kangaroos. Preliminary attempts at short-term preservation showed that the motility of kangaroo spermatozoa could be adequately stored for 24 h and that the addition of egg yolk to the semen diluent was beneficial for improving the rate of sperm movement.