Toxoplasma gondii, an obligate intracellular parasite, is the aetiological agent of toxoplasmosis, a disease that affects approximately 25% - 30% of the world's population. At present, no safe and effective vaccine exists for the prevention of toxoplasmosis. Current treatment options for toxoplasmosis are active only against tachyzoites and may also cause bone marrow toxicity. To contribute to the global search for novel agents for the treatment of toxoplasmosis, we herein report the in vitro activities of previously synthesised benzyltriazole derivatives. The effects of these compounds against T. gondii in vitro were evaluated by using a expressing green fluorescent protein (GFP) type I strain parasite (RH-GFP) and a type II cyst-forming strain of parasite (PruΔku80Δhxgprt). The frontline antitubercular drug isoniazid, designated as Frans J. Smit -isoniazid (FJS-INH), was also included in the screening as a preliminary test in view of future repurposing of this agent. Of the compounds screened, FJS-302, FJS-303, FJS-403 and FJS-INH demonstrated > 80% parasite growth inhibition with IC50 values of 5.6 µg/mL, 6.8 µg/µL, 7.0 µg/mL and 19.8 µg/mL, respectively. FJS-302, FJS-303 and FJS-403 inhibited parasite invasion and replication, whereas, sulphadiazine (SFZ), the positive control, was only effective against parasite replication. In addition, SFZ induced bradyzoite differentiation in vitro, whilst FJS-302, FJS-303 and FJS-403 did not increase the bradyzoite number. These results indicate that FJS-302, FJS-303 and FJS-403 have the potential to act as a viable source of antiparasitic therapeutic agents.

There are limited data on the efficacy of antiparasitic treatments and husbandry methods to control nematode infections in captive populations of African green monkeys (AGMs), Chlorocebus sabaeus. In faecal egg count (FEC) tests, 10 of the 11 (91%) adult male AGMs captured from the large feral population on the island of St Kitts had evidence of nematode infections, mostly Capillaria (8/11, 73%), Trichuris trichiura (7/11, 64%) and strongylid species (7/11, 64%) specifically (hookworm and Trichostrongylus, 50/50), but also Strongyloides fuelleborni (1/11, 9%). When kept in individual cages with cleaning and feeding regimens to prevent reinfections and treated concurrently with ivermectin (300 µg/kg, given subcutaneously) and albendazole (10 mg/kg, given orally) daily for 3 days, 60% (6/10) of the AGMs were negative at a follow-up FEC at 3 months and by FEC and necropsy at the end of the study 5-8 months later. One monkey appeared to have been reinfected with T. trichiura after being negative by FEC at 3 months post-treatment. Four AGMs were positive for T. trichiura at the 3 month FEC follow-up but were negative at the end of the study after one further treatment regimen. Although initially being cleared of Capillaria following treatment, three AGMs were found to be infected at the end of the study. The ivermectin and albendazole treatment regimen coupled with good husbandry practices to prevent reinfections effectively controlled nematode infections in captive AGMs.

In this study, the serological surveillance of Rift Valley Fever virus (RVFV) in southern Egypt was carried out for 460 serum samples collected from domestic animals (unvaccinated), including cattle, sheep, goat, camel and donkey reared in three different provinces (Qena, Luxor and Aswan). Enzyme linked immunosorbent assay (ELISA) was used to detect RVFV antibodies. The results showed that 97 out of 460 animals were positive by using blocking ELISA. The percentage of RVFV infection in cattle, sheep, goat, camel and donkey was 5.55%, 65.21%, 14.44%, 20.65% and 0%, respectively. Geographical distribution and breeding system were taken into consideration for RVFV infection in these animals. The most prevalent type of infection was identified in intensive breeding farms systems (27.63%), and then in individual breeding systems (11.68%). Qena had a higher infection rate of RVFV (23.55%), in comparison to Aswan and Luxor (20.65% and 14.14%, respectively). Marked seroprevalence recorded in this study indicates a high incidence of infection in sheep (65.21%) and camel (20.65%); this necessitates the application of more effective strategies to control these types of infections in Egypt. This study provides a concise picture about the RVFV disease in southern Egypt. We need more similar studies targeted to clarify the reliable epidemiological status of RVFV disease in southern Egypt and other localities.

OBJECTIVE To evaluate surfactant protein D (SP-D) concentrations in serum and bronchoalveolar lavage fluid (BALF) from young healthy horses on pasture or housed in a typical barn. ANIMALS 20 young healthy horses. PROCEDURES Horses were randomly assigned to 1 of 2 groups (pasture, n = 10; barn, 10), and serum and BALF samples were collected for SP-D determination at baseline (all horses on pasture) and 2 weeks and 4 weeks after the barn group of horses was relocated from the pasture to the barn. Other evaluations included physical and tracheoscopic examinations. Findings were compared within and between groups. RESULTS Physical and tracheoscopic examinations, CBC, and serum biochemical analysis did not reveal evidence of respiratory disease, and no significant differences were present within and between groups. Serum SP-D concentrations did not significantly differ within and between groups, but BALF SP-D concentrations were significantly lower for the barn group at 2 weeks but not at 4 weeks, compared with baseline. The BALF SP-D concentration-to-BALF total protein concentration ratio was < 1.5 and did not significantly differ within and between groups. CONCLUSIONS AND CLINICAL RELEVANCE A mild decrease was evident in the concentration of SP-D in the BALF collected from young healthy horses after 2 weeks of exposure to a barn environment. The clinical importance of this finding remains to be determined.

The aim of this study was to evaluate the occurrence of Shiga toxin (stx)-producing Escherichia coli (STEC) in diarrheic newborn calves, as well as the resistance profile of this microorganism against antimicrobials routinely used in veterinary therapy. The antimicrobial profile of Eugenia uniflora against E. coli clinical isolates was also analyzed. Specimens from the recto-anal junction mucosa were investigated by using chromogenic medium and identification of E. coli was done using microbiological methods (Gram staining, indole test, methyl red test, Voges-Proskauer test, citrate test, urease test, and hydrogen sulfide test). The stx1 and stx2 genes corresponding to the STEC pathotype were evaluated by using polymerase chain reaction and electrophoresis. The susceptibility profile to antimicrobial agents commonly used in veterinary therapeutic practice and the antimicrobial effect of lyophilized hydroalcoholic extract of E. uniflora L. leaves against E. coli clinical isolates were evaluated by disk diffusion and microdilution methods. Shiga toxin-positive E. coli was identified in 45% of diarrheic newborn calves (stx1 = 23.2%, stx2 = 4.0%, stx1 + stx2 = 18.2%). The frequency of stx-positive E. coli in the bacterial population was equal to 17.0% (168/990 clinical isolates): 97 (9.8%) stx1-positive E. coli, 12 (1.2%) stx2-positive E. coli, and 59 (6.0%) stx1 + stx2-positive E. coli isolates. All stx-positive E. coli analyzed showed resistance to multiple drugs, that is, from 4 to 10 antimicrobials per clinical isolate (streptomycin, tetracycline, cephalothin, ampicillin, sulfamethoxazole + trimethoprim, nitrofurantoin and nalidixic acid, ciprofloxacin, gentamicin, and chloramphenicol). Effective management measures should be implemented, including clinical and laboratory monitoring, in order to promote animal and worker health and welfare, prevent and control the spread of diseases, and ensure effective treatment of infectious diseases. The E. uniflora L. leaves showed inhibition of microbial growth based on the diameter of halos, ranging from 7.9 to 8.0 mm and 9.9 to 10.1 mm for concentrations of 50 and 150 mg/mL, respectively. This plant displayed bacteriostatic action and a minimum inhibitory concentration of 12.5 mg/mL for all clinical isolates. Its clinical or synergistic effects with antimicrobial agents must be determined from clinical and preclinical trials.

Changes in immune factors expressed by milk somatic cells from Holstein cows with hypocalcemia after calving were investigated in this study. Fourteen multiparous Holstein cows after their 3rd or 4th calving in one farm were used. The cows were divided into 2 groups: 7 cows needing treatment due to onset of hypocalcemia (hypocalcemia group; age = 5.53 ± 0.27 years, parity = 3.14 ± 0.14) and 7 cows without health problems (control group; age = 5.88 ± 0.31 years, parity = 3.57 ± 0.26). Milk samples were collected aseptically using a cannula and mRNA of immune factors expressed by milk somatic cells were analyzed. Milk samples (50 mL) were collected from the right rear mammary gland of cows before milking at day 1 and weeks 1, 2, 4, and 8 after calving. All milk samples showed a negative reaction to the California Mastitis Test. Levels of relative interleukin (IL)-6 and cathelicidin in the hypocalcemia group were lower than those in the control group in weeks 1 to 8. A significant difference in relative IL-6 levels was found in week 4 (P < 0.05). These results suggest that levels of IL-6 expressed by milk somatic cells may be affected by hypocalcemia in dairy cows.

OBJECTIVE To evaluate physical compatibility of small animal (SAE) and large animal (LAE) injectable formulations of enrofloxacin with select IV fluids and drugs. SAMPLE 162 admixtures containing SAE or LAE with saline (0.9% NaCl) solution, lactated Ringer solution (LRS), Plasma-Lyte A (PLA), 6% hydroxyethylstarch 130/0.4 (HES), metoclopramide, or ampicillin-sulbactam. PROCEDURES In the first of 2 simultaneously conducted experiments, admixtures containing enrofloxacin (10 mg/kg) and a volume of IV fluid that would be administered over a 20-minute period when dosed at the maintenance infusion rate (40 mL/kg/d for saline solution, LRS, and PLA and 20 mL/kg/d for HES) were created. In the second experiment, enrofloxacin (10 mg/kg) was admixed with saline solution (40 mL/kg/d) and metoclopramide (2 mg/kg/d) or ampicillin-sulbactam (30 mg/kg). In both experiments, admixture components were infused into a flask over 20 minutes assuming patient weights of 5, 10, and 20 kg. Admixtures were created by use of undiluted SAE and SAE diluted 1:1 with saline solution and undiluted LAE and LAE diluted 1:1 and 1:10 with saline solution. Admixtures were assessed for physical incompatibility at 0, 15, 30, and 60 minutes after completion of mixing. Physical incompatibility was defined as gross precipitation, cloudiness, Tyndall effect, or change in turbidity. RESULTS Admixtures containing undiluted SAE or LAE were physically incompatible with saline solution, PLA, LRS, and HES. Because saline solution was used to dilute SAE and LAE, all admixtures containing diluted SAE or LAE were also physically incompatible. Physical compatibility of enrofloxacin with metoclopramide or ampicillin-sulbactam could not be assessed because those admixtures also contained saline solution. CONCLUSIONS AND CLINICAL RELEVANCE Enrofloxacin was physically incompatible with all tested solutions.

OBJECTIVE To evaluate synoviocentesis of the equine forelimb digital flexor tendon sheath (DFTS) via a basilar sesamoidean approach (BSA) or distal approach (DA). ANIMALS 21 healthy adult horses without DFTS-related lameness. PROCEDURES The forelimbs of each horse underwent the BSA or DA (21 limbs/approach) performed by 1 individual. The volume of synovial fluid (SF) aspirated, time from skin puncture to collection of SF, and number of attempts to place a needle in the DFTS were compared between approaches. RESULTS An SF sample was successfully aspirated from 16 of 21 (76%) limbs with the BSA and 20 of 21 (95%) limbs with the DA. For the BSA and DA, the number of attempts to obtain SF was 2 and 1, respectively; the median volume of SF obtained was 0.4 and 0.7 mL, respectively; and the median time to SF collection was 17.91 and 18.48 seconds, respectively. Between the approaches, the number of limbs with SF successfully aspirated and number of attempts to collect SF differed significantly, whereas the volume of SF aspirated and time to SF collection did not. CONCLUSIONS AND CLINICAL RELEVANCE Regarding SF collection from forelimb DFTSs in horses without DFTS-related disease, use of the DA had a greater success rate with fewer attempts, compared with findings for the BSA, which may reflect the relative ease of identifying anatomic landmarks for the DA. Results suggested that a DA for DFTS synoviocentesis in horses appears efficient and effective and may minimize limb trauma by requiring fewer attempts for SF sample collection, compared with a BSA.

The objective of this preliminary study was to identify genomic regions that may predispose Gordon setters from the United Kingdom to familial protein-losing enteropathy (PLE) at a young age. A total of 106 related Gordon setters was used, including 6 affected dogs from an affected litter, 6 case controls from the same litter, 10 related/affected dogs, and 84 related/unaffected dogs. Genomic DNA was collected from each Gordon setter and extracted from buccal mucosal swabs. Genotyping of affected and unaffected dogs was carried out using the Canine Illumina HD SNP array and data generated were analyzed with PLINK software, using fixation index (Fst) and runs of homozygosity (ROH) methods. Pairwise Fst analyses between the affected and unaffected Gordon setter dogs identified various regions of differentiation on chromosomes 10, 18, 21, and 23 that contained several important genes. These regions revealed 5 candidate genes, including RARB, TTC7A, SOCS5, PIGF, and RHOD, that are associated with human inflammatory bowel disease (IBD) and could potentially be associated with PLE in Gordon setters. Run of homozygosity (ROH) analyses revealed additional unique regions on chromosomes 15 and 17. These regions contained genes SYT1, UCN, and FNDC that could also be potential candidates for PLE in Gordon setters. The biological functions of the identified genes provided initial insights into the pathophysiology of PLE. Further large-scale studies are warranted to investigate the possible causality of these genomic regions and any possible genetic markers that could be used in predicting susceptibility to PLE syndrome.

OBJECTIVE To compare progesterone (P4) concentrations measured with surface plasmon field-enhanced fluorescence spectroscopy (SPFS) and chemiluminescence immunoassay (CLIA) in serum and plasma samples of client-owned bitches of various ages and breeds and to determine reference ranges for P4 concentrations at various stages of the estrous cycle. SAMPLES 102 serum samples and 104 plasma samples. PROCEDURES In experiment 1, 1 aliquot each of serum and plasma was analyzed for P4 concentration by use of SPFS incorporated in a veterinary-specific point-of-care immunologic analyzer and CLIA. In experiment 2, serum collected from bitches in various stages of the estrous cycle was analyzed for P4 concentration by use of SPFS to establish reference ranges for each stage. RESULTS In experiment 1, P4 concentrations measured by SPFS and CLIA were highly correlated (serum, r = 0.966; plasma, r = 0.968). In experiment 2, ranges of serum basal (proestrous) P4 concentrations (n = 114) and P4 concentrations at the estimated time of ovulation (76), during pregnancy or diestrus (107), and during the prepartum period (50) measured with SPFS were 0.42 to 1.46 ng/mL, 3.69 to 7.85 ng/mL, 11.73 to 28.24 ng/mL, and 1.54 to 3.22 ng/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Because serum and plasma P4 concentrations measured with SPFS were highly correlated with those measured with CLIA and ranges of serum P4 concentrations measured with SPFS for each of phase of the estrous cycle were well-defined for the large sample size, veterinarians may be able to accurately use this veterinary-specific point-of-care immunologic analyzer with SPFS methodology to determine P4 concentrations of bitches in their daily practice.