Toxoplasma gondii infection in domestic ducks, free-range and caged chickens in southern China

Toxoplasma gondii is widely distributed in humans and other animals including domestic poultry throughout the world, but little is known of the prevalence of T. gondii in chickens and ducks in People's Republic of China. In the present study, antibodies to T. gondii were investigated in 349 domestic ducks (Anas spp.), 361 free-range, and 244 caged chickens (Gallus domesticus) raised in commercial flocks in Southern China's Guangdong Province using the modified agglutination test (MAT). Antibodies to T. gondii (MAT titer of 1:5 or higher) were found in 56 (16%) of 349 ducks, 41 (11.4%) of 361 free-range, and 10 (4.1%) of 244 caged chickens. The results indicate soil contamination due to T. gondii oocysts because free-range chickens feed from the ground, and suggest that the meat from the domestic poultry may be an important source for human infection by T. gondii in People's Republic of China.     

Seroprevalence of Toxoplasma gondii infection in sheep and alpacas (Llama pacos) in Chile

Serum samples from 408 sheep from different regions of Chile and 447 alpacas (Llama pacos) from the north of the country were tested for Toxoplasma gondii antibodies. The indirect haemagglutination test (IHAT) was used in both species and the indirect immunofluorescence test (IIFT) was also used on the sheep samples in order to compare the performance of the tests in that species. In both tests, titers ≥1:16 were considered diagnostically significant. Sera from 49 sheep (12%) were positive to T. gondii antibodies by the IHAT. When using the IIFT, 114 sheep sera (28%) were positive. The different results obtained in sheep sera between the tests were significant (p<0.0001). No differences were observed between geographical locations or sex of the sampled sheep regarding serological detection of T. gondii antibodies in sheep. As expected, adult sheep showed higher T. gondii reactivity than young sheep (p=0.0008). The corrected prevalence of toxoplasmosis in alpaca was 16.3% (32 positive out of 447). The rather low prevalence in alpacas may be associated with their extensive management as well as the extreme climatic conditions of The Andes which apparently would not be favorable for the transmission of the parasite.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Mossoró, Rio Grande do Norte, Brazil

Toxoplasma gondii is an obligate intracellular parasite with a variety of hosts, responsible for reproductive problems and economic losses in sheep flocks. Neospora caninum was recently identified and its clinical presentation in sheep is similar to that of toxoplasmosis, which can cause repeated abortions, though less frequently in this species. In order to confirm the prevalence of these agents in the city of Mossoró, Rio Grande do Norte, Brazil, 409 serum samples from adult sheep (364 females and 45 males) were tested by the indirect immunofluorescence antibody test, using cut-off point at a dilution of 1:64 and 1:50 for T. gondii and N. caninum, respectively. From the 35 properties examined, 23 (65.7%) had at least one seropositive animal for T. gondii and six (17.1%) for N. caninum. The prevalence of seropositive animals for T. gondii was 20.7% and for N. caninum 1.8%. There was no association between the presence of the agent’s antibody and gender, reports of reproductive problems and presence of dogs and/or cats in the properties. T. gondii is well distributed and N. caninum has low prevalence in sheep and in the properties of the studied region.     

Mycoplasma synoviae invades non-phagocytic chicken cells in vitro

Mycoplasma synoviae and Mycoplasma gallisepticum are major poultry pathogens, but their strains differ significantly in invasiveness and pathogenicity. Recent studies have demonstrated that M. gallisepticum invades chicken erythrocytes (CER) and chicken embryonic fibroblasts. The aim of this study was to determine whether M. synoviae also invades chicken cells. Using the gentamicin invasion assay, relative invasion frequency (RIF) of four M. synoviae strains was determined for CER, chicken embryonic cell line (CEC-32) and/or primary chicken chondrocytes (CCH). All tested strains of M. synoviae were capable of invading chicken cells within 24 h after infection. The type strain WVU 1853 showed significantly higher invasiveness in CER (RIF 7.5 ± 1.5%) and CEC-32 (RIF 7.0 ± 0.3%) than field strain ULB 02/T6 and M. gallisepticum strain R_low. Surprisingly, WVU 1853, which is capable of causing synovitis and arthritis in chickens, was less invasive for CCH with a RIF (1.2 ± 0.3%) similar to that of R_low (1.1 ± 0.1%). This is the first study documenting the invasiveness of M. synoviae strains for non-phagocytic chicken cells.     

Distribution of cysts and tachyczoites in calves and pregnant cows inoculated with toxoplasma gondii oocysts

Sixteen calves and 6 cows were each inoculated with 100 000 infective oocytes of the GT-1 strain of Toxoplasma gondii. Cattle were necropsied between 3 and 287 days post-inoculation (DPI) and their tissues were inoculated into mice or fed to Toxoplasma-free cats for the detection of Toxoplasma in bovine tissues. Ten to 10 000-fold more T. gondii were recovered from small intestine and mesenteric lymph nodes of calves at 3 and 6 DPI than from lungs and liver, and the number of T. gondii in bovine tissues was reduced 1000-fold between 6 and 8 DPI. By using the pepsin digestion technique, or feeding tissues to Toxoplasma-free cats, it was demonstrated that T. gondii encysted in bovine tissues as early as 11 DPI and persisted as late as 287 DPI. More Toxoplasma gondii cysts occurred in livers than in any other bovine tissue. Of the 6 cows inoculated at 95–155 days after breeding, 5 delivered normal calves and T. gondii was isolated from only one of these calves. One cow was barren. Toxoplasma gondii was not isolated either by mouse inoculation or by feeding cats tissues from 2 cows killed 132 and 190 DPI. Toxoplasma gondii was not isolated in mice inoculated with tissues of cows killed 98 and 109 DPI, but cats fed on bovine tissues shed T. gondii oocysts. The organism, however, was isolated in mice inoculated with the mesenteric lymph nodes of 1 of the 2 cows killed 162 and 168 DPI, and from the small intestine of the other. Cats fed tissues of these cows later shed T. gondii oocysts.     

First identification of Sarcocystis tenella (Railliet, 1886) Moulé, 1886 (Protozoa: Apicomplexa) by PCR in naturally infected sheep from Brazil

Sarcocystis tenella is a dog–sheep protozoan parasite, causing a widespread enzootic muscle parasitosis and neurological disease mainly in lambs. This parasite is pathogenic to sheep and important to the economical production of sheep. The present study was initially aimed to determine Toxoplasma gondii infection and the occurrence of co-infection with other Apicomplexa parasites in 602 Brazilian sheep. Twenty of these sheep were positive with antibodies to T. gondii by MAT and IFAT-IgG tests, positive with PCR-RFLP genotyping at multiple loci, and parasites were isolated from mice infected with sheep tissue samples. Two additional sheep born in Brazil, a 2-year-old female Polwarth (Ideal) sheep, a breed originated from Australia (#1), and a 1-year-old male Corriedale sheep, a breed originated from New Zealand and Australia (#2) were positive to T. gondii antibodies by serum tests, and PCR, but negative for bioassay in mice. In genotyping at 12 loci, sheep #1 sample and #2 presented positive results only for some markers. PCR-RFLP of 18S ribosomal RNA (18S rRNA) was performed in all 22 animals to identify the possibility of co-infection of T. gondii with other Apicomplexa parasites, such as S. tenella, Neospora caninum and Hammondia hammondi, resulting in a T. gondii profile for the first 20 animals and a unique genotyping profile for sheep #1 and #2, identical to S. tenella. The 18S rRNA PCR products (310 bp) were sequenced and blasted to GenBank database at NCBI. Both samples were identical to S. tenella 18S rRNA gene (GenBank accession number L24383-1). These results suggest the existence of co-infection of S. tenella with T. gondii in ewes from Brazil.     

弓形虫细胞核因子3基因的克隆及序列分析

为预测弓形虫细胞核因子3(TgNF3)基因作为抗弓形虫疫苗候选抗原的可能性,本试验以弓形虫RH株的总RNA为模板,应用逆转录-聚合酶链式反应(RT-PCR)扩增TgNF3基因并连接至pMD18-T载体。筛选阳性克隆后,测序鉴定。将测序结果正确的核苷酸序列翻译成氨基酸序列,运用生物信息学软件分析TgNF3蛋白的结构,预测抗原表位。结果显示,TgNF3基因长约950bp。TgNF3蛋白中包含1个跨膜结构域,9个α-螺旋区,4个β-折叠区,8个亲水区域和10个柔性区域,并预测含有9个线性B细胞抗原表位。结果表明,TgNF3蛋白可能具有良好的免疫原性,为研制以TgNF3为抗原的弓形虫亚单位疫苗及表位疫苗奠定了基础。     

Responses to condensed tannins of flowering sulla (Hedysarum coronarium L.) grazed by dairy sheep: Part 1: Effects on feeding behaviour, intake, diet digestibility and performance

The concentration of condensed tannins (CT) in sulla (Hedysarum coronarium L.) is moderate and overall regarded as beneficial. However, the intake of this forage can reduce diet digestibility, particularly during flowering phase. An experiment was run to assess the effect of CT on feeding behaviour, intake, diet digestibility and performance of dairy sheep rotationally grazing sulla at flowering phase. Twenty-four late-lactating sheep were blocked in two homogeneous groups and submitted to the following treatments: i) twice daily drenching with 200 g/day of a 50% w/v water solution of an anti-tannic substance, polyethylene glycol, group PEG; ii) twice daily drenching with 200 g/day of water, group CON (Control group). All the sheep rotationally grazed as a flock two sulla plots from April to June (8 weeks in total). Sward height, herbage mass, botanical and chemical composition of the herbage were measured at the beginning and the end of each grazing period. The feeding behaviour (3 sheep per group) was continuously monitored for 24 h in 6 weeks using the IGER behaviour recorders. Herbage DM intake (DMI), dietary DM digestibility (DMD) and apparent CP digestibility (CPD) were estimated on 8 sheep per group by the n-alkane method. On average, PEG group had longer total grazing (503 ± 12 vs 460 ± 12 min, P < 0.05) and eating time (425 ± 13 vs 391 ± 13 min, P < 0.07) than CON group. Moreover PEG group showed shorter inter-meal intervals (41 ± 3 vs 52 ± 3, min, P < 0.05) and higher number of daily meals than CT-exposed group (24 ± 1 vs 19 ± 1 min, P < 0.01). The herbage DMI was not affected by the treatment whereas DMD (74.60 ± 3.48 vs 58.30 ± 3.01%), and CPD (60.14 ± 4.83 vs 38.21 ± 4.83%) were both increased by PEG administration (P < 0.05) confirming the negative effect of sulla CT on these variables. Milk yield tended to be higher in PEG than CON (1331 ± 45 vs 1205 ± 59 ml, P < 0.11). Milk protein content was similar between groups while milk fat content was higher in CON than PEG ewes (6.61 ± 0.15 vs 6.11 ± 0.15%, P < 0.05), being the reverse true for milk urea (46.04 ± 1.27 vs 53.04 ± 0.76%, P < 0.001). To conclude, this experiment shows that when sulla is grazed at flowering as monoculture, dietary CT can exert negative effects on DM and CP apparent digestibility, in this study partially counterbalanced by a better metabolic utilisation of the nutrients up-taken.     

Effects of Saccharomyces cerevisiae on ruminal pH and microbial fermentation in dairy cows: Yeast supplementation on rumen fermentation

An experiment was conducted with eight ruminally-cannulated cows using a crossover design with 2 periods to determine the effects of yeast supplementation on rumen fermentation. Holstein dairy cows in late lactation were either supplemented with 0.5 g/hd/d of Saccharomyces cerevisiae, an active dry yeast (CNCM-1077, Levucell SC20 (r) SC, Lallemand Animal Nutrition) or not supplemented (control). A basal diet consisting of 60% forage and 40% concentrate (DM basis) was fed once daily to both groups of cows throughout the entire experiment. Ruminal pH was measured continuously every 22 min using a pH probe that was placed in the ventral rumen sac for 6 consecutive days. Volatile fatty acid and ammonia N concentrations in the rumen were measured on days 5 or 6 of the 12-d period for each cow and DM intake was monitored throughout the experiment. Data were analyzed using a mixed-effects model with repeated measures. There were no differences in dry matter intake between treatments. Mean ruminal pH was greater (P < 0.05) when yeast was supplemented (6.53 ± 0.07) compared with the control (6.32 ± 0.07). Average maximum and minimum ruminal pH were also greater (P < 0.05) when yeast was supplemented (7.01 ± 0.09 and 5.97 ± 0.08, respectively) compared with the control (6.80 ± 0.09 and 5.69 ± 0.09, respectively). Time spent under the subacute acidosis threshold, pH less than 5.6, was lower (P < 0.05) with yeast supplementation compared with control cows. No difference was observed for ruminal ammonia N concentrations (mean = 14.0 ± 1.2 mg/dL) between treatments. Total VFA concentration (mM) in the rumen tended to be lower (P = 0.10) in the yeast-supplemented cows (107.3 ± 6.35) than in the control cows (122.4 ± 6.35), which could be related to the greater pH observed with yeast supplementation. Supplementing dairy cows with active dry yeast in the current experiment increased the mean, minimum and maximum ruminal pH; decreased time spent in subacute rumen acidosis, and tended to decrease total VFA concentration in the rumen compared with control cows.     

Life-history parameters of Culicoides (Avaritia) imicola Kieffer in the laboratory at different rearing temperatures

This laboratory study investigates the sub-adult developmental cycle of field collected Culicoides (Avaritia) imicola Kieffer (Diptera; Ceratopogonidae). The period required from blood-feeding field-collected females to the production of progeny adults occupied 34–56 days at 20 °C, 15–21 days at 25 °C and 11–16 days at 28 °C, demonstrating clear temperature dependence. When reared at 28 °C, C. imicola demonstrated higher variability in fecundity (between 2.4 and 20.6 eggs/female) and lower hatching rates (50.0–62.2%), although larval survival rates to pupation were low at all temperatures (20–30%). Similarly, the mean emergence rate from pupae was the highest at lower temperatures. These results highlight the difficulty in establishing and maintaining a laboratory colony of this species from field-collected material and results are discussed in reference to future research directions that may aid this process.     

Factors affecting seroprevalence of Toxoplasma gondii in the endangered Iberian lynx (Lynx pardinus)

Wild felids are considered important in maintaining the sylvatic cycle of Toxoplasma gondii. Although, T. gondii antibodies have been reported in several species of wild felids, little is known of the epidemiology and risk factors associated with T. gondii infection in wild cats. The Iberian lynx (Lynx pardinus) is the most endangered felid species in the world. In the present study, seroprevalence and associated risk factors for T. gondii infection in a large population of Iberian lynx in Spain were determined. Serum samples from 129 Iberian lynx collected from 2005 to 2009 and 85 wild rabbits (Oryctolagus cuniculus), sharing the habitat with the Iberian lynx, were tested for antibodies to T. gondii by the modified agglutination test (MAT) using a cut-off value of 1:25. Antibodies to T. gondii were found in 81 of 129 (62.8%) Iberian lynx. Seroprevalence to T. gondii in Iberian lynx significantly increased with age (P < 0.001). T. gondii seroprevalences were similar in free-ranging (66.7% of 93) and wild-caught captive lynx (69% of 84) but significantly lower in captive-born lynx (22.5% of 40). Seroprevalence was higher in lynx with concurrent Cytauxzoonfelis (88% of 25) but not with concurrent Feline Leukemia Virus (FeLV) infection (53.8% of 13). There were no significant differences in seroprevalence between sexes, geographic region and year of sample collection (2005–2009). Oocysts of T. gondii were not detected microscopically in fecal samples from 58 lynx. Wild rabbits are considered the most important food for the lynx. Antibodies to T. gondii were found in 14 (11.9%) of 85 rabbits tested. The present results indicate that T. gondii infection is widespread in the two areas where Iberian lynx survive in Spain. The fact that four captive-born lynx seroconverted was indication of contact with T. gondii also in the Captive Breeding Centers, hence, control measures to prevent T. gondii infection would be necessary in these centers.     

The use of a semiochemical bait to enhance exposure of Amblyomma variegatum (Acari: Ixodidae) to Metarhizium anisopliae (Ascomycota: Hypocreales)

Experiments were conducted to explore the use of a semiochemical bait to enhance exposure of Amblyomma variegatum Fabricius (Acari: Ixodidae) to different formulations of the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorok. (Ascomycota: Hypocreales). Initially, the relative efficacies of attraction-aggregation-attachment pheromone (AAAP), made up of o-nitrophenol, methyl salicylate and nonanoic acid in the ratio 2:1:8, 1-octen-3-ol and butyric acid, were evaluated in an olfactometer. Only AAAP and 1-octen-3-ol were found to elicit attractive responses to the tick. Simultaneous release of 1-octen-3-ol and AAAP together with CO₂ from a trap in semifield plots attracted up to 94.0 ± 6% of adult ticks from a distance of 6 m, and up to 24.0 ± 5.1% from 8 m. Formulations of M. anisopliae (dry powder, oil, and emulsifiable) applied within the trap baited with AAAP, 1-octen-3-ol and CO₂ resulted in high levels of contamination of the ticks attracted to the traps. However, 48 h after autoinoculation, 89.1 and 33.3% of conidia were lost in dry powder and oil formulations, respectively. Emulsifiable formulation showed least loss of propagules (17.1%). Samples of ticks attracted to the baited traps were transferred to plastic basins containing grass and maintained for 5 weeks. The experiment was conducted in rainy and dry seasons. Emulsifiable formulation gave the highest relative tick reduction in both seasons: 54.7 and 46.5% in rainy and dry seasons, respectively, followed by oil formulation (32.0 and 23.8%) and powder formulation (38.0 and 24.4%).     

Prevalence of Toxoplasma gondii antibodies in stray cats in Sari, northern Iran

Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that can excrete environmentally resistant oocysts. T.gondii is a major zoonotic agent which infects up to one-third of the world population. Toxoplasmosis in neonates and immunocompromised patients can lead to severe disease and death. A cross- sectional parasitological and serological survey with latex agglutination test (LAT) to detect anti-T. gondii antibodies was conducted on 100 serum samples collected from stray cats in five urban areas of Sari, Northern Iran, from April to November 2004. Classification by age, sex, weight, season and region was made. Results analyzed according to specific variables. The overall prevalence of T. gondii IgG antibodies (LAT titre ≥1:1) were found in 40 of 100(40%) of stray cats, with regional variations. Overall 16 of 100(16%) of stray cats had diagnostically significant antibody titres (LAT ≥ 1:64). Prevalence was significantly higher in adult cats (1.5–3.0 kg, 54.5% of 66) than in juvenile cats and kittens (≤1.4 kg, 11.8% of 34) and higher in female stray cats (44.4% of 72) than in male stray cats (28.6% of 28). Toxoplasma seroprevalence was highest in the season of spring (22.4%). There was a significant difference in the prevalence of infection relative to host age and weight (P < 0.05). No significant difference was found between the prevalence of infection relative to host gender, urban sites and season (P > 0.05). Prevalence of T. gondii oocyst was also analyzed by a routine coprological method in 100 cats. T. gondii oocysts were not found in any faecal samples analyzed. Only 2 out of 100 smear preparations of intestinal mucosa showed trophozoites of T. gondii.     

The protective effect of a Toxoplasma gondii SAG1 plasmid DNA vaccine in mice is enhanced with IL-18

More effective vaccines against Toxoplasma gondii may contribute to the control of this pathogen that has major veterinary and public health significance. In this study, two recombinant plasmids pcDNA/TgSAG1 and pVAX/mIL-18 containing T. gondii SAG1 (TgSAG1) and murine cytokine interleukin-18 (IL-18) were evaluated for their ability to protect mice against T. gondii challenge. Mice were given two intramuscular immunizations 3 weeks apart, and challenged with T. gondii 3 weeks later. All animals vaccinated with pcDNA/TgSAG1 alone or with pVAX/mIL-18 developed specific anti-TLA (T. gondii lysate antigen) antibodies and specific lymphocyte proliferative responses. Co-injection of pVAX/mIL-18 significantly increased the production of IFN-γ and IL-2. Further, challenge experiments showed that co-immunization with pVAX/mIL-18 significantly (P < 0.05) increased the survival rate (60%), compared with pcDNA/TgSAG1 alone (40%). Therefore, codelivery of the IL-18-secreting plasmid potentiates the induction and maintenance of the type 1 helper T-cell immune response and may be a potent strategy for enhancing the protective efficacy of vaccines against T. gondii.     

Antibody-ELISA for Trypanosoma evansi: Application in a serological survey of dairy cattle, Thailand, and validation of a locally produced antigen

Trypanosoma evansi is generally considered a mild pathogen in bovines. However, in Asia, acute and chronic signs have been observed in cattle, with high levels of parasitaemia, abortion and death. Investigations in Asian cattle are needed to better understand this epidemiological situation. To generate comparable data at a regional level, development and standardization of an antibody-enzyme linked immunosorbent assay for T. evansi (ELISA/T. evansi) was initiated and applied in an epidemiological survey carried out in dairy cattle in Thailand. A batch of 1979 samples was collected from dairy farms located throughout the country's four regions. Soluble T. evansi antigens initially produced in France were also produced in Thailand for comparison and technology transfer. Screening of 500 samples allowed us to identify reference samples and to determine the cut-off value of the ELISA. Seropositive animals – some of them confirmed by PCR – were found in the four regions, in 12 out of 13 provinces, in 22 out of 31 districts, in 56 farms out of 222 (25%, 95%CI ± 6%) and in 163 animals out of 1979 (8.2, 95%CI ± 1.2%). Estimated seroprevalence in 35 farms ranged between 1% and 30%, and in 21 farms it was >30%. Approximately 25% of survey cattle were exposed to the infection, in various situations. A sub-sample of 160 sera was tested on both antigens. Wilcoxon's (Z = 1.24; p = 0.22) and McNemars's tests (CHI2 = 3.55; p = 0.09) did not show any significant differences, showing that the locally produced antigen is suitable for further evaluation in the surrounding countries. Use of this standardized serological method will broaden knowledge of the prevalence and impact of the disease at the regional level in South-East Asia. Further validation of this ELISA will be necessary in other host species such as buffalo, horse and pig.     

Dietary supplementation with Acanthopanax senticosus extract enhances gut health in weanling piglets

The present study was conducted to investigate the effects of Acanthopanax senticosus extract (ASE) as a dietary additive on gut health in weanling piglets by examining diarrhea frequency, intestinal microbiota and morphology. A total of 96 Duroc× (Landrace × Yorkshire) piglets weaned at 21 days of age with an average initial body weight (BW) of 5.6 ± 0.4 kg were randomly assigned to 3 treatment groups with 4 duplicates of 8 piglets each. The piglets were fed basal diet to which had been added 0 or 1 g/kg of ASE, or 0.7 g/kg antibiotics, respectively. Fecal consistence was monitored twice daily and the frequency of diarrhea was calculated. On day 21 after the initiation of supplementation, 8 piglets were randomly selected from each treatment group (2 piglets per pen) and slaughtered. The jejunum, ileum, colon and cecum were then excised and fixed in 10% neutral formalin solution to determine villus height and crypt depth, after their contents were collected to determine microbiota. The results showed that dietary supplementation with ASE increased (P < 0.05) the density of bacterial populations that co-migrated with Lactobacillus amylovorus, Lactobacillus salivarius, Bacillus subtilis, and Clostridium lituseburens, but decreased (P < 0.05) those co-migrating with Staphylococcus aureus, Salmonella typhimurium, Ruminococcus forques, and E. coli O157:H7 in the PCR-DGGE profiling analysis when compared with the control group. The villus height of the duodenum, jejunum and ileum increased (P < 0.05) by 14.8, 13.7 and 10.0%, while the crypt depth decreased (P < 0.05) by 17.9, 9.1 and 12.1%, respectively, in response to dietary ASE supplementation. Additionally dietary supplementation with ASE or an antibiotic decreased (P < 0.05) the frequency of diarrhea by 55.6 and 52.2%, respectively, compared with the control group. In conclusion, these findings suggest that dietary supplementation with ASE could regulate the microbiota composition and maintain a normal morphology of gut mucosa in weanling piglets, thereby decreasing diarrhea that resulted from weaning stress.     

Effects of synbiotic-based Bifidobacterium animalis in female rats experimentally infected with Toxoplasma gondii

The aim of this study was to assess the effects of a synbiotic composed of Bifidobacterium animalis and fructooligosaccharides on female rats infected with Toxoplasma gondii. Female Wistar rats, treated or not with dexamethasone, were daily supplemented with synbiotics for 21 days. After 15 days of supplementation, the rats were orally infected with 10⁴T. gondii bradyzoites. Blood samples were collected to measure the levels of IFN-γ, IL-10 and T. gondii antibodies. All synbiotic-supplemented rats survived until the end of the experiment; however, non-supplemented dexamethasone-treated rats died between the fifth and the eighth days after T. gondii infection. Dexamethasone-treated rats supplemented with synbiotics (P < 0.05) were capable of synthesizing IFN-γ, and this immunological response was essential to ensure their survival. In addition, brain cysts were found in one rat not supplemented with synbiotics. Results suggest that the synbiotic composed of B. animalis and fructooligosaccharides may be beneficial to toxoplasmosis control.     

Pharmacokinetics of marbofloxacin after a single oral dose to loggerhead sea turtles (Caretta caretta)

The single-dose disposition kinetics of marbofloxacin (MBX) were determined in clinically healthy loggerhead sea turtles (n = 5) after oral (PO) administration of 2 mg kg⁻¹ bodyweight. Marbofloxacin plasma concentrations were determined by DAD–HPLC (LOD/LOQ 0.015/0.05 μg ml⁻¹). Data were subjected to non-compartmental analysis. Following PO administration, marbofloxacin achieved maximum plasma concentrations of 11.66 ± 2.53 mg L⁻¹ at 15.00 ± 3.00 h. The absence of general adverse reactions in the turtles of the study, and the favourable pharmacokinetic properties (long half-life and high maximum plasma concentration) of MBX administered PO at the single-dose of 2 mg kg⁻¹ suggest the possibility of its safe and effective clinical use in loggerhead sea turtles.     

Diet composition, herbage intake and digestibility in Inner Mongolian Cashmere goats grazing on native Leymus chinensis plant communities

The botanical composition, intake and digestibility of the diet selected by Inner Mongolian Cashmere goats grazing on a native Leymus chinensis plant community were measured in two grazing periods (June and August) using the n-alkane markers. In each grazing period, 48 Inner Mongolian Cashmere goats (24 wethers and 24 does with an average live weight of 22.1 ± 1.2 kg and 19.6 ± 0.8 kg, respectively) were divided in two groups (G1: grazing group; G2: cage-fed group) of 24 animals each. Based on the observation of G1 grazing behaviour, G2 goats were fed on herbage obtained by a simulative method and were housed in metabolism cages to determine the alkane faecal recoveries. Diet composition was estimated by comparing the odd-chain n-alkane pattern (C₂₅–C₃₅) of the selected plant species with the n-alkane faecal concentrations, corrected for their incomplete faecal recovery. The alkane pair C₃₂:C₃₃ and C₃₆ alkane were used to estimate intake and diet digestibility, respectively. The results showed that the increase of herbage availability from June to August, resulted in higher (P < 0.05) DM, CP, ME, ADF and NDF intake of both wethers and does, with no evident changes in ADF and NDF intake in does., The intake of nutrients differed significantly between wethers and does in both experimental periods, except for NDF in EP1 and ME in EP2. Differences between grazing seasons and goats' sex in nutrients digestibility were not observed. Diet composition indicated seasonal differences, being Sonchus brachyotus, Artemisia frigida and Phragmites communis the main components of the diet in June. However, Suaeda corniculata, Chloris virgata, Xanthium sibiricum and Echinochloa crusgalli became the main diet components in August. Moreover, the sex affected significantly the diet choices in both grazing periods.     

A novel enhanced dot blot immunoassay using colorimetric biosensor for detection of Toxoplasma gondii infection

This study reports development of a novel point of care assay, namely an enhanced immuno-dot blot assay, for discrimination of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies. This method has been designed based on formation of a sandwich complex between a gold nanoprobe (chitosan gold nanoparticle-anti-human IgG or anti-IgM) and anti- Toxoplasma lysate antigen (TLA) which holds anti-TLA antibodies, either IgG or IgM. Briefly, anti-human IgG or anti-IgM antibody was conjugated to chitosan gold nanoparticles via glutaraldehyde chemistry. Then, lysate antigen was immobilized on the surface of nitrocellulose membrane, which followed by addition of the sera sample and gold nanoprobes. The positive signals were readily detectable via observation with naked eye. This positive color change was further intensified via gold enhancement chemistry. The intensity of biosensor signal was proportional to the concentration of active antibodies on the surface of nanoparticles, titer of T. gondii antibodies in the sera samples and concentration of Toxoplasma lysate antigen coated on the nitrocellulose membrane. A minimum concentration to use the antibodies for conjugation, to detect titer of Toxoplasma IgG and IgM antibodies, and the concentration of TLA coated in nitrocellulose membrane were 0.5 mg/mL, 2 IU/mL, 10 IU/mL, and 20 μg/mL, respectively. This enhanced immuno-dot blot assay offers a simple diagnostic technique without expensive equipment requirement for distinguishing of anti- T. gondii IgM and IgG antibodies in field conditions, pregnant women, and immunocompromised patients.