The prevalence and pathogenicity of Chorioptes bovis (Hering, 1845) and Psoroptes cuniculi (Delafond, 1859) (Acari: Psoroptidae) infestations in feral goats in New Zealand

Between October 1976 and January 1980 a total of 368 feral goats was examined in New Zealand for the presence of Chorioptes bovis and 434 for the presence of Psoroptes cuniculi. The seasonal pattern of C. bovis infestation in feral goats was similar to that seen in sheep and cattle. The prevalence of infestation reached 100% in July and August (winter) and declined to 27% in February and March (summer). Fewer goats were infested with P. cuniculi and prevalence of infestation reached a maximum of 41% in July. No goats were found infested in the December and January samples. Chorioptes bovis infestation was independent of the age and sex of hosts. Infestation with P. cuniculi was independent of the sex of the hosts, but older goats were more frequently infested (16%) than younger goats (8.6%). Unilateral infestation with P. cuniculi was more common (74.5%) than bilateral infestation. No severe lesions were associated with C. bovis, but 21.3% of goats with P. cuniculi had the external auditory meatus blocked by waxy material and 12.8% had scabby encrustations on the ears. The importance of goats as a possible source of infestation to domestic animals is discussed.     

Excretory–secretory antigens: A suitable candidate for immunization against ocular toxoplasmosis in a murine model

Toxoplasmosis, responsible for ocular impairment, is caused by Toxoplasma gondii. We investigated the effect of Toxoplasma excretory–secretory antigens (ESA) on parasite load and distribution in the eye tissue of a murine model. Case and control groups were immunized with ESA and PBS, respectively. Two weeks after the second immunization, the mice were challenged intraperitoneally with virulent RH strain of Toxoplasma; eye tissue samples of both groups were collected daily (days 1, 2, 3, and the last day before death). Parasite load was determined using real-time quantitative PCR targeted at the B1 gene. Compared to the control group, infected mice that received ESA vaccine presented a considerable decrease in parasite load in the eye tissue, demonstrating the effect of ESA on parasite load and distribution. Diminution of parasite load in mouse eye tissue indicated that ESA might help control disease-related complications and could be a valuable immunization candidate against ocular toxoplasmosis.     

Molecular detection and genotype identification of E. cuniculi from pet rabbits

Encephalitozoon cuniculi is a microsporidian which is frequently reported from rabbits. This microorganism can either ravage rabbit farms or transmit to humans from pet rabbits. This study aimed to investigate the prevalence and the genotype distribution of E. cuniculi among pet rabbits. In this study urine samples were collected from 50 pet rabbits, aged 2 months to 3 years, admitted to teaching veterinary hospital. Four races Lop, Dutch, Mix, and Angora were screened for E. cuniculi. The clinical symptoms were recorded and total DNA was extracted from urine samples. E. cuniculi was identified using amplification of the small subunit ribosomal RNA (ssu rRNA) gene and its genotypes were characterized using PCR/sequencing of the polar tube protein (PTP) gene. Phylogenetic tree was drawn to confirm the characterized genotypes. Out of 50 samples, 41 (82 %) of rabbits were asymptomatic, while nine (18 %) had at least one of symptoms including head-tilt, circling, and ataxia. A statistical correlation was seen between mean age + SD and symptoms (P-value = 0.039). The presence of E. cuniculi was confirmed in 16/50 (32 %) rabbits and all of them were identified as the genotype I. Our findings represented no consistency between E. cuniculi PCR – positive and the presence of symptoms (P-value = 0.318). Our results showed positive correlation between symptoms and age; however, the lack of correlation between PCR results with age may signify the latent infection in younger rabbits. All identified E. cuniculi were the genotype I, which is reported from rabbits and humans, highlighting the zoonotic concern for this genotype, particularly among subjects who keep pet rabbits.     

ANTIMICROBIAL SUSCEPTIBILITY PATTERNS OF BIOFILM FORMING STAPHYLOCOCCUS AUREUS ISOLATED FROM PIGEON EXTERNAL OCULAR INFECTIONS

Pathogenic microorganisms are commonly associated with external ocular infections in birds. Pathogen virulence factors as well as reduced host defenses resulting from poor living conditions, nutrition, genetics, physiology, hygiene, fever, and age may increase host susceptibility. Staphylococcus species are bacteria known to serve as opportunistic pathogens in eye infections. The changing profile of microorganisms involved in ocular infections and the emergence of acquired microbial resistance dictate the need for investigative studies regarding bacterial profiles and antimicrobial susceptibility patterns for external ocular infections. The aim of this study was to determine the prevalence of Staphylococcus aureus ocular infections in pigeons and to evaluate their biofilm production ability and antibiotic resistance patterns. Twenty pigeons with confirmed eye infections were included in this project. Conjunctival specimens were collected with swabs presoaked in sterile normal saline. Bacterial growth was identified by standard laboratory procedures and susceptibility testing of the isolates was performed by the Kirby-Bauer method. The ability of the isolates to form a biofilm was also assessed using the microtiter plate method. Of the 20 specimens processed, 20% of the pigeons had staphylococcal eye infections. The resistance pattern of these isolates showed that Staphylococcus spp. from pigeon samples were resistant to tetracycline (100%), erythromycin (100%), azithromycin (100%), nalidixic acid (100%), and cefazolin (50%). All of the Staphylococcus spp. isolated from the pigeons were susceptible to gentamycin and furazolidone. The results of the biofilm detection test showed that 75% of the isolates were biofilm producers. In conclusion, biofilm forming S. aureus with multidrug resistance patterns were the most prevalent bacteria isolated from the pigeons examined in this research study.     

Encephalitozoon cuniculi infection in rabbits

The microsporidian parasite Encephalitozoon cuniculi commonly infects rabbits. Most infections are initially asymptomatic, but for reasons yet to be explained, many rabbits subsequently develop disease as a result of infection with this organism. Three common forms of this disease are recognized, and they can occur individually or in combination. The ocular form is associated with cataracts and when there is extensive damage to the lens, uveitis. The neurological form can vary from a mild change in the rabbit’s behavior to severe vestibular disease. The signs associated with the renal form of the disease are those of chronic progressive renal disease. Definitive diagnosis of encephalitozoonosis in the rabbit is difficult. Animals with encephalitozoonosis are expected to be seropositive, but many apparently healthy rabbits are also seropositive, so this assay is not specific and its results must be considered in the light of other diagnostic findings. The absence of antibody, however, should cause the practitioner to consider other differentials. Drugs proven to be efficacious for E. cuniculi infections include albendazole and fenbendazole. Supportive care and treatment with antiinflammatory medications may also be necessary in some forms of encephalitozoonosis. Lens removal or removal of the lens contents is indicated in some rabbits with E. cuniculi-induced ocular disease.     

Mycobacterium genavense Infection in a Domestic Ferret (Mustela putorius furo)

Mycobacterium genavense infection was diagnosed in an adult ferret with ptosis of the left eye, a proliferative lesion of the conjunctiva of the nictitating membrane, conjunctival swelling, and tumefaction of the periorbital tissues with a watery ocular discharge and the presence of a retrobulbar mass. The diagnosis was based on characteristic cytology of the retrobulbar mass and left mandibular lymph node that revealed granulomatous inflammation. Ziehl-Neelsen staining showed the presence of positive acid-fast bacilli in the cytoplasm of the macrophages. The diagnosis was confirmed by sequence analysis of the 16S rRNA gene amplified by using a multiplex polymerase chain reaction from a fresh lymph node biopsy. Therapy with marbofloxacin, rifampicin, and clarithromycin was recommended for 6 months and after this period, the veterinarian who was treating the ferret reported the disappearance of clinical signs. Six months after the end of the antibiotic treatment, the symptoms described previously reoccurred. Confirmatory laboratory tests were not performed but a recurrence of M genavense infection was suspected and the veterinarian, in agreement with the owner, euthanized the ferret.     

Detection of Encephalitozoon cuniculi in the feline cataractous lens

Purpose Identification of Encephalitozoon cuniculi (E. cuniculi) as a possible causative agent for cataracts and uveitis in cats. Methods Within a 12‐month study period, cats that were presented with focal anterior cortical or mature cataract and secondary uveitis underwent a complete ophthalmic examination, complete blood count, serum biochemistry, serologic tests for E. cuniculi and tests for feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), feline leukemia virus (FeLV) and Toxoplasma gondii (T. gondii). PCR for DNA detection of E. cuniculi and T. gondii as well as cytologic examination of aqueous humor after paracentesis and phacoemulsified lens material were also performed. In addition histopathologic examination of the resected anterior lens capsule and attached lens epithelial cells was performed. Serologic testing for antibodies against E. cuniculi was also performed in 100 ophthalmologically healthy cats. Results Eleven (19 eyes) European shorthair cats with a median age of 3.5 years were included. Nine of 11 cats had bilateral cataracts, with 12/19 eyes having focal anterior cortical cataracts and 7/19 eyes having mature cataracts. In 14/19 eyes anterior uveitis was present. All cats had a positive antibody titer (1:80–1:10 000) for E. cuniculi. Encephalitozoon cuniculi DNA was detected by PCR and sequencing in 18/19 lenses and in 10/19 aqueous samples. Five tentative positive results were detected by cytologic examination. Spores were detected in 15/19 samples of lens material with histopathologic staining. Only 2/100 ophthalmologically healthy cats showed a positive antibody titer for E. cuniculi. Conclusion Encephalitozoon cuniculi is a cause of focal anterior cortical cataract and anterior uveitis in cats.     

Salmonella enterica serotype typhimurium and S. Stanley differ in genomic evolutionary patterns and early immune responses in human THP-1 cell line and CD14+ monocytes

Salmonella Typhimurium and S. Stanley are the most prevalent serogroup B serovars to infect humans in Taiwan. The aim was to determine possible factors to influence the prevalence between S. Typhimurium and S. Stanley. Genotypes were determined by pulsed field gel electrophoresis (PFGE) analysis and the intracellular survival, phagocytosis, reactive oxygen species (ROS) production of human monocyte THP-1 cell and tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6), and IL-1βexpression in peripheral blood CD14⁺ cells after infection were analyzed. 182 S. Stanley was clonal disseminated with main pulsotypes 2 from 2004 to 2007. Overall S. Typhimurium evolved more genotypes, while S. Stanley conserved in genotypes. Human blood CD14⁺ monocytes expressed TNF-α, IL-6 and IL-1β differently among serovars and bacterial conditions (live vs. killed). Live S. Stanley and S. Typhimurium suppressed the TNF-α and IL-6 expression compared to killed bacteria. However, live S. Typhimurium stimulated more IL-1β expression than the killed bacteria, but S. Stanley expressed similar IL-1β levels in both conditions. Furthermore, S. Stanley and S. Typhimurium differed in intracellular survival in the THP-1 cells, an early decrease for S. Stanley, not for S. Typhimurium. Additionally, higher reactive oxygen species (ROS) production in THP-1 cells was found agsinst S. Stanley infection, not found in S. Typhimurium. However, some isolates of S. Stanley could recover from early loss to become more in the monocytes than S. Typhimurium. Difference in phagocytized number, intracellular survival, ROS production and IL-1β expression may contribute to prevalence different between two serovars.     

Evaluation of the live vaccine efficacy of virulence plasmid-cured,and phoP- or aroA-deficient Salmonella enterica serovar Typhimurium in mice

We evaluated the protective efficacy of 94-kb virulence plasmid-cured, and phoP- or aroA-deficient strains of Salmonella enterica serovar Typhimurium (ΔphoP or ΔaroA S. Typhimurium) as oral vaccine candidates in BALB/c mice. Two weeks after the completion of 3 oral immunizations with 1 × 10⁸ colony-forming units (CFU) of virulence plasmid-cured, and ΔphoP or ΔaroA S. Typhimurium at 10-day intervals, S. Typhimurium lipopolysaccharide (LPS)-specific mucosal secretory immunoglobulin A (s-IgA) antibody titers were detected in the cecal homogenate, bile and lung lavage fluid, but not in the intestinal lavage fluid. In addition, the increases in S. Typhimurium LPS-specific immunoglobulin G (IgG) and IgA antibody titers in the serum were also observed 2 weeks after completing 3 oral immunizations with virulence plasmid-cured, and ΔphoP or ΔaroA S. Typhimurium. The series of 3 oral immunizations protected the mice against an oral challenge with 5 × 10⁸ CFU of the virulent strain of S. Typhimurium, suggesting that both the virulence plasmid-cured, and ΔphoP and ΔaroA S. Typhimurium strains are promising candidates for safe and effective live S. Typhimurium vaccines.     

First insight into Encephalitozoon cuniculi infection in laboratory and pet rabbits in Iran

Encephalitozoon cuniculi infects a wide variety of domestic and wild mammalian species including humans. Although the infection status has been studied in laboratory and pet rabbits worldwide, there is shortage of information regarding the disease in Iran. In the present study, the occurrence of infection in brains of 117 asymptomatic rabbits from six breeding and experimental units with highest population of rabbit colonies in the country (n = 60) as well as pet rabbits of pet stores in two cities (n = 57) were examined by nested-PCR. Histological sections of brains and kidneys were also studied by light microscopy. PCR results revealed that 3.3% of laboratory rabbits (2/60) and 59.6% of pet rabbits (34/57) harboured E. cuniculi in their brains. Histopathology on the other hand showed spores of the parasite in kidney and brain of one and kidney of another pet rabbit. As encephalitozoonosis may interfere with results of experiments performed on laboratory rabbits, routine screenings for identification and culling of infected animals is recommended. Furthermore, infected companion rabbits can transmit E. cuniculi to people in close contact with them, therefore, improving public knowledge of this zoonotic infection is suggested.     

A multiplex real-time PCR for differential detection and quantification of Salmonella spp., Salmonella enterica serovar Typhimurium and Enteritidis in meats

Salmonella (S.) Typhimurium and S. Enteritidis are the major causative agents of food-borne illnesses worldwide. Currently, a rapid detection system using multiplex real-time polymerase chain reaction (PCR) has been applied for other food-borne pathogens such as Escherichia coli, Staphylococcus aureus and Streptococcus spp. A multiplex real-time PCR was developed for the simultaneous detection of Salmonella spp., especially S. Typhimurium and S. Enteritidis, in beef and pork. For the specific and sensitive multiplex real-time PCR, three representative primers and probes were designed based on sequence data from Genbank. Among the three DNA extraction methods (boiling, alkaline lysis, and QIAamp DNA Mini Kit), the QIAamp DNA Mini Kit was the most sensitive in this study. The optimized multiplex real-time PCR was applied to artificially inoculated beef or pork. The detection sensitivity of the multiplex real-time PCR was increased. The specificity of the multiplex real-time PCR assay, using 128 pure-cultured bacteria including 110 Salmonella isolates and 18 non-Salmonella isolates, was 100%, 100% and 99.1% for Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. The sensitivity was 100%, 100% and 91.7% for Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. The multiplex real-time PCR assay developed in this study could detect up to 0.54 ± 0.09 and 0.65 ± 0.07 log₁₀ CFU/ml for S. Typhimurium and S. Enteritidis for beef, 1.45 ± 0.21 and 1.65 ± 0.07 log₁₀ CFU/ml for S. Typhimurium and S. Enteritidis for pork, respectively, with all conditions optimized. Our results indicated that the multiplex real-time PCR assay developed in this study could sensitively detect Salmonella spp. and specifically differentiate S. Typhimurium from S. Enteritidis in meats.     

Serological survey of Encephalitozoon cuniculi in farm rabbits in Italy

Rabbit sera (n = 1600) from 40 commercial farms were submitted to a serological screening for Encephalitozoon cuniculi by an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA test). Antibodies anti-Encephalitozoon cuniculi were found in 505/1600 (31.6%) sera analysed, and all the farms (100%) resulted positive. Rabbits older than 4 months showed a significantly higher seropositivity for E. cuniculi (chi-squared test: p < 0.0001) than rabbits under 4 months, E. cuniculi sero-prevalence showed an increasing trend in rabbits within the farm along with the increase in the “number of rabbits on the farm”; however, this trend was not significant (Spearman’s correlation: p = 0.073).The findings of the present study confirm that rabbit is the main reservoir of E. cuniculi; they are of epidemiological relevance and immediate public health importance because of the recognized infectivity in humans by the microsporidium.     

Acuaroid nematodes in the common kestrel (Falco tinnunculus) in the south of Spain

The prevalence, intensity and abundance of acuaroid nematodes were determined in the common kestrel (Falco tinnunculus) in Andalusia, Spain. Acuaroid nematodes were present in 26/41 (63.4%) of birds examined. The most common species belonged to the genus Synhimantus subgenus Synhimantus (56%): S. (S.) laticeps (36.5%), S. (S.) robertdollfusi (24.3%) and a single specimen of a third, unknown, Synhimantus (S.) spp., unlike any other described previously (2.4%). Other species identified were Synhimantus (Dispharynx) spp. (2.4%), S. (D.) nasuta (4.8%), Desportesius spinulatus (9.7%) and Skrjabinoclava spp. (2.4%). This is the first record of these three species in F. tinnunculus, but the latter two are considered to be accidental parasites in birds of prey.     

Encephalitozoon cuniculi–Associated Equine Encephalitis: A Case Report

A case of encephalitis of unknown origin in the horse was investigated. Postmortem examination findings revealed a nonsuppurative granulomatous meningoencephalitis in the right hemisphere of the cerebral cortex. Testing for West Nile virus, equine herpes virus, equine infectious anemia, Toxoplasma gondii, Neospora caninum, and Sarcocystis neurona were negative. The horse had a titer for Encephalitozoon cuniculi, and sections from the affected area of the brain tested positive for the organism using both polymerase chain reaction (PCR) and immunohistochemistry. Amplicons generated using PCR were sequenced, and E. cuniculi genotype II was identified. This is the first case of E. cuniculi genotype II associated with encephalitis in the horse.     

Attenuated Salmonella choleraesuis-mediated RNAi targeted to conserved regions against foot-and-mouth disease virus in guinea pigs and swine

In this study, specific sequences within three genes (3D, VP4 and 2B) of the foot-and-mouth disease virus (FMDV) genome were determined to be effective RNAi targets. These sequences are highly conserved among different serotype viruses based on sequence analysis. Small interfering RNA (siRNA)-expressing plasmids (p3D-NT19, p3D-NT56, pVP4-NT19, pVP4-NT65 and p2B-NT25) were constructed to express siRNA targeting 3D, VP4 and 2B, respectively. The antiviral potential of these siRNA for various FMDV isolates was investigated in baby hamster kidney (BHK-21) cells and suckling mice. The results show that these siRNA inhibited virus yield 10- to 300-fold for different FMDV isolates of serotype O and serotype Asia I at 48 h post infection in BHK-21 cells compared to control cells. In suckling mice, p3D-NT56 and p2B-NT25 delayed the death of mice. Twenty percent to 40% of the animals that received a single siRNA dose survived 5 days post infection with serotype O or serotype Asia I. We used an attenuated Salmonella choleraesuis (C500) vaccine strain, to carry the plasmid that expresses siRNA directed against the polymerase gene 3D (p3D-NT56) of FMDV. We used guinea pigs to evaluate the inhibitory effects of recombinant S. cho (p3D-NT56/S. cho) on FMDV infection. The results show that 80% of guinea pigs inoculated with 10⁹ CFU of p3D-NT56/S. cho and challenged 36 h later with 50 ID₅₀ of homologous FMDV were protected. We also measured the antiviral activity of p3D-NT56/S. cho in swine. The results indicate that 100% of the animals treated with 5 × 10⁹ CFU of p3D-NT56/S. cho were protected in 9 days.     

Effect of substitution of concentrate mix with Sesbania sesban on feed intake, digestibility, body weight change, and carcass parameters of Arsi-Bale sheep fed a basal diet of native grass hay

An experiment was conducted to assess the effect of substitution of concentrate mix with Sesbania sesban on feed intake, digestibility, average daily gain (ADG), and carcass parameters of Arsi-Bale sheep. The experiment employed 25 male sheep with mean (±standard error) initial body live weight (BLW) of 19.1?±?0.09 kg. The experiment consisted of 7 days of digestibility and 90 days of feeding trials followed by carcass evaluation. The experiment employed a randomized complete block design with five treatments and five blocks. Treatments comprised of grass hay alone fed ad libitum (GHA; control), GHA?+?100 % concentrate mix (CM) consisting of wheat bran and noug seed cake at a ratio of 2:1 (0 S. sesban), GHA?+?67 % CM?+?33 % S. sesban (33 S. sesban), GHA?+?33 % CM?+?67 % S. sesban (67 S. sesban), and GHA?+?100 % S. sesban (100 S. sesban). Total dry matter intake (DMI) was higher (p?<?0.001) for sheep in 0 S. sesban–100 S. sesban (800–821 g/day) compared to sheep in control (611 g/day). However, the effect of S. sesban inclusion (0 S. sesban–100 S. sesban) on total DMI was quadratic, and DMI declined after 67 S. sesban. Digestibility of DM, organic matter (p?<?0.01), and crude protein were higher (p?<?0.001) in supplemented group compared to the control. ADG, feed conversion efficiency (ADG/DMI), slaughter BLW, hot carcass weight, and total edible offals were higher (p?<?0.05–0.001) for sheep in 0 S. sesban–100 S. sesban than those in control. Increased level of S. sesban inclusion, in general, reduced growth and carcass parameters in this study. However, there was no difference between 0 S. sesban and 33 S. sesban in most parameters studied. Thus, it can be concluded that S. sesban could substitute a concentrate when it accounted for up to 33 % of the mix.     

Characterization of a Streptococcus equi ssp. equi Isolate From a Strangles Outbreak in Thailand

Strangles, which is caused by Streptococcus equi ssp. equi, is one of the major infectious respiratory diseases in horses. Knowledge of isolates from different areas of the world is important for investigating the different strains of the disease. In contrast to many other countries, currently little is known about S. equi ssp. equi isolates in Thailand. In 2014, a farm in Thailand imported 20 horses from Europe. Approximately 1 month after arrival, 50% of the horses had developed pyrexia, mucopurulent nasal discharge, and abscesses of the mandibular lymph nodes. Nasal swabs of mucopurulent discharge were sent to a diagnostic laboratory, and two isolates of S. equi ssp. equi were identified. One of the isolates was further characterized using seM gene polymerase chain reaction and sequence analysis. The seM sequence was then compared to the database of PubMLST-seM. It was found to contain SeM allele 48, an allele isolated from horses in the United Kingdom in 2006 and 2010. This result demonstrates the usefulness of SeM allele identification as a tool for investigating the source of related strains and for the epidemiologic study of strangles. To the best of the authors' knowledge, this is the first report of the identification of an SeM allele of the S. equi ssp. equi isolate in Thailand.