Efficacy of dietary live or heat-killed Bacillus subtilis in goldfish (Carassius auratus) infected with Ichthyophthirius multifiliis.

BACKGROUND: The beneficial effects of Bacillus subtilis on growth, immune response, and disease resistance against various diseases in different fish species have been proved. However, there are no data concerning this probiotic effect on skin mucosal immunity in fish infected with Ichthyophthirius multifiliis (Ich). Ich has a high mortality rate in both edible and ornamental fish and consequently is concerned with heavy economic losses. OBJECTIVES: Thus, we assessed the efficacy of live and heat-killed B. subtilis on skin immunity and histopathology in goldfish (Carassius auratus) infected with Ich. METHODS: Goldfish (144 fish, 2.38 g average weight) were stocked in nine glass tanks each in three replicates. Fish were fed 109 CFU g-1 live or heat-killed B. subtilis for 80 days. RESULTS: Probiotic administration in both viable and non-viable forms could enhance the growth performance in goldfish. Probiotic therapy also reduced the density of the parasite and histopathological level on skin and gill tissues of the treated fish. Real-time polymerase chain reaction analysis showed a higher expression of lysozyme and tumour necrosis factor-α in the treated groups compared to the control group. CONCLUSIONS: These data demonstrated the beneficial effect of B. subtilis as probiotic and paraprobiotic on growth performance and disease resistance to Ich infestation in goldfish.

Natural infection with Toxoplasma gondii, Neospora caninum and Sarcocystis species in domestic pigeons (Columba livia domestica) in Iran.

Pigeons are common birds around the world and may act as intermediate hosts of the tissue cyst-forming apicomplexan parasites Toxoplasma gondii, Neospora caninum and Sacrocystis spp. This study aimed to provide an overview on the prevalence of and exposure to these parasites in Iranian domestic rock pigeon (Columba livia domestica) through molecular, serological and histopathological examination. Blood and tissue samples (i.e., brain, heart, gizzard, neck, thigh, and pectoral muscles) were taken from 100 pigeons. Sera were screened by agglutination tests for detection of anti- T. gondii and N. caninum antibodies, genomic DNA from tissue samples were assessed by respective species-specific PCRs, and histopathological examination of tissues was carried out. A seroprevalence of 45 % to anti-T. gondii and 35 % to anti-N. caninum IgG was recorded. PCR detected T. gondii DNA in 28 pigeons. Sacrocystis spp. was detected in one animal, but sequencing of the 28 S rRNA gene product did not reveal the identity of the species. Histopathology revealed myocarditis, myositis, and gliosis in the heart, skeletal muscles, and brain, respectively. No Sarcocystis tissue-cysts were detected, but T. gondii tissue cyst-like structures in the brain (i.e., 4 %) and heart (i.e., 3 %) were found by histology. Data reported herein demonstrate that pigeons from Iran are infected with tissue cyst-forming apicomplexans, particularly T. gondii. Since domestic pigeons are in close contact with human populations, and consumption of their meat and egg is popular in different societies, control strategies for minimizing the risk of infection in both pigeons and humans are suggested.

The Protective Effect of Artemisia spicigera Ethanolic Extract against Cryptosporidium parvum Infection in Immunosuppressed Mice.

BACKGROUND: A specific treatment has not yet developed for cryptosporidiosis, and some of the used drugs had side effects in immunodeficient patients. The goal of an appropriate remedy is to remove symptoms and improve immune responses in hosts. The current study was designed to evaluate the therapeutic efficacy of Artemisia spicigera ethanolic extract in experimentally infected immunosuppressed mice. METHODS: Thirty six NMRI mice, 4-6 wk old, were randomly divided into six equal groups. C1: uninfected, treated control; C2: infected, untreated control; T1, T2, T3, and P: infected, treated with 0.2, 2, and 20 mg/ml extract, and 5mg/ml paromomycin, respectively. Mice were experimentally infected by oral administration of 104 oocysts/animal of Cryptosporidium parvum and treated orally for eight days per 12h, starting 12h before experimental infection. The presence of oocyst shedding, weight gain/loss, and the histopathology of ileum sections were examined. RESULTS: Results revealed that oocyst shedding was significantly (P<0.05) reduced in treatment groups. There was no significant difference between the mean of weight gain/loss in the infected control and treated groups. Histopathological analysis of ileum sections further supported the parasitological findings. CONCLUSION: Artemisia spicigera had acceptable efficacy as a therapeutic agent for cryptosporidiosis.

First Survey on the Presence and Distribution of Oxytetracycline-Resistance Genes in Anaplasma Species.

INTRODUCTION: Anaplasma sp. is an obligatory intracellular Gram-negative tick-transmitted bacterial pathogen of humans and animals. Oxytetracycline and chlortetracycline are the drugs of choice for treating domestic animals with acute anaplasmosis. Lack of documented information about oxytetracycline resistance in Anaplasma species in the world was the scope of this study to screen by PCR for the detection of the oxytetracycline-resistance genes in Anaplasma species from infected cattle and sheep in the Northwest and Southwest of Iran. MATERIALS AND METHODS: Total of 100 cattle and sheep blood samples collected from 2 provinces in the Northwest and 1 province in the Southwest of Iran were tested microscopically by the Giemsa staining examination and confirmed by PCR. Then the presence of two different oxytetracycline-resistance genes (otrA, and otrB) was detected by PCR in positive samples. RESULTS: The results showed that 60% of Anaplasma-infected samples were identified to have an otrA-resistance gene, and 26.67% had an otrB-resistance gene. The coexistence of two oxytetracycline-resistance determinants was encountered in 13.33% of the isolates. The significant difference in the frequency of otr genes was found among three Anaplasma species (A. marginale, A. centrale and A. ovis), and among three studied regions in Iran (p < 0.05). The identified sequences were submitted to the GenBank and deposited under accession numbers MN880729 and MN895439 for otrB and otrA genes. CONCLUSION: This study, for the first time, indicated the oxytetracycline-resistance genes in the three most prevalent Anaplasma species in ruminants. This finding helps to select an appropriate treatment strategy for eradication of anaplasmosis.

Molecular study and genotyping of Cryptosporidium baileyi and Cryptosporidium parvum from free-range and commercial broiler chickens in Guilan province, Iran.

Cryptosporidiosis acutely impacts the digestive and/or respiratory tract of the birds in many species of various orders. More importantly, it is also well known as a significant zoonotic disease, which can lead to diarrhea in humans and livestock. Regarding increasing demand for free-range products and increasing the number of free-range poultry farms, the present paper evaluated histopathological and molecular detection of Cryptosporidium baileyi and Cryptosporidium parvum in free-range and commercial broiler chickens in the north part of Iran. For this purpose, 100 fecal and tissue samples of the chickens in Guilan province were collected. After microscopic examination using Ziehl-Neelsen staining, molecular analyses of the fecal samples were processed by Nested-PCR targeting the 18S rRNA gene followed by sequencing of the amplicons and phylogenetic analyses. Eventually, the tissue samples were studied for histological lesions. Findings demonstrated the presence of Cryptosporidium baileyi and Cryptosporidium parvum in 6 % and 2 % of fecal samples, respectively. This is the first identification of C.parvum in avian hosts in Iran, and for the first time, C.baileyi and C.parvum are shown in native free-range chickens in Iran. All of the PCR positive birds with clinical symptoms showed gross lesions of respiratory infections. There was no significant difference between infection rate in free-range and commercial broiler chickens; however, the infection rate was significantly higher in chickens <25 days old. To conclude, we present here a notable Cryptosporidium infection rate in the free-range chicks in Iran, which notify the role of this host as a reservoir and should be more noted due to the economic and zoonotic importance.

Comparative survey on scolicidal effects of selenium and silver nanoparticles on protoscolices of hydatid cyst.

Hydatidosis is a zoonotic diseases which affects human and livestock animals. The surgical treatment of hydatid diseases in man is still the most effective approach, but spillage of protoscolices is the risk of surgery. Therefore, it is essential to develop alternative approaches. Injection of protoscolicidal agents into the cysts preoperatively is an alternative method.The aim of the present study was the determination and comparison of lethal effects of silver nanoparticles (Ag-NPs) and selenium nanoparticles (Se-NPs) on protoscolices of hydatid cyst. Livers and lungs of sheep, which was naturally infected by hydatid cyst were collected from Tabriz slaughterhouse and transferred to the parasitology laboratory. Initially, the surface of cysts was disinfected by 70% Alcohol. Then, 25 ml of each cyst fluid were aspirated by sterile syringe and transferred to Erlenmeyer flask. After 30 minutes, the supernatant was discarded and protoscolices were washed with phosphate-buffered saline and tested by 1% eosin to assess their viability. Next, the samples of protoscolices, which were over 90%, were selected and the effect of 50, 125, 250 and 500 (µg/ml) of Se-NPs and Ag-NPs in the exposure times of 10, 20, 30 and 60 min were evaluated. Data were analysed by SAS software and analyzed by Duncan's comparison test. The significant differences were considered to be p<0.0001.The difference between the scolicidal effects of Se-NPs was statistically significant for all examined concentrations and exposure times compared to the control group (p<0.0001). On the other hand, the scolicidal effect of Ag-NPs in 10 and 20 min exposer was not significant in comparison to the control group. Overall scolicidal effect of Se-NPs was higher than Ag-NPs and the difference was significant (p<0.0001).The results of the present research indicated that it is possible to use Se-NPs as an effective scolicidal treatment. It is suggested the study of employing this compound in vivo or in combination with other procedures in treating hydatid cyst in further studies.

Computational, experimental details, and biological raw data accompanying the publication: "The synthesis and characterization of a nanomagnetite with potent antibacterial activity and low mammalian toxicity".

This data file includes experimental details on how to make uncoated iron oxide nanoparticles using a green electrochemical method. It provides the raw data on the antibacterial activity of one of these formulations, and the full computational data and methodology used to generate that data, of several different magnetite clusters of specific spin multiplicities for 4, 5, 7 and 9 iron atom magnetite clusters. This data will assist other researchers wishing to replicate or expand on these results for the investigation and use of nanomagnetite for antibacterial applications.

A study on rate of infestation to Sarcocystis cysts in supplied raw hamburgers.

This study was carried on for determination of presence of Sarcocystis cysts in raw hamburgers in Tabriz North West of Iran. Ninety-six samples of industrial (70 % meat content) and traditional (30 % meat content) hamburgers (80 samples industrial and 16 samples traditional) were obtained from retail fast food stores. The samples were examined by gross examination, and microscopic examination methods consist impression smear and peptic digestion. Macroscopic cysts did not observed in any of the samples in gross examination. Microscopic study showed that from 96 samples 54 (56.25 %) samples were infected by at least one bradyzoites of Sarcocystis. From 54 infected samples, 45 industrial hamburgers and nine traditional hamburgers samples were infected. Statistical analysis showed that there was not significant differences between industrial and traditional hamburgers in infection to Sarcocystis. Infestation of hamburgers to Sarcocystis in summer was higher than other seasons but this difference was not significant. In Iran, beef meat is used for preparation of 70 % of hamburger and infestation of cattle to sarcocystosis was reported in many investigations in Iran. With regard to the high prevalence of Sarcocystis infection in meat products such as hamburgers in this study, it is strongly recommended to avoid eating raw or under-cooked hamburgers or keep them at freezing temperature for at least 3-5 days.

Application of recombinant Cryptosporidium parvum P23 for isolation and prevention.

Cryptosporidium parvum is a coccidian protozoan that causes diarrhea in immunocompromised humans and newborn animals. Billions of oocysts of C. parvum can be released from the infected calves and can contaminate the environment. The severity of the disease depends on the immunological status of the individual. Oocysts of Cryptosporidium are extremely resistant to many environmental stresses, and no effective disinfectant and curative agent against this organism is available. In our study, recombinant C. parvum P23 was prepared for application in the isolation and prevention of cryptosporidiosis. P23 is a glycoprotein that belongs to a family of protein of 23-27 kDa and is believed to be expressed in the different life stages of the parasite. Immunostaining analysis using the indirect fluorescent antibody test showed that P23 could be recognized on the surface of the oocysts. The antibody prepared in rabbit against P23 was bound to Sepharose 4B and used for the isolation of oocysts. The results showed that the prepared column was able to bind specifically only the oocysts. The effect of specific recombinant C. parvum IgY antibody against infection with C. parvum was examined in a mouse model. For this aim, purified egg yolk antibody prepared from immunized hens was used to analyze the protective effect of recombinant P23 specific antibody in immunosuppressed adult mice. The results showed more than 70% reduction in oocyst shedding after challenge with 1 × 10(4) oocysts. These results support previous studies of other investigators regarding the protective effect of P23 as an antigen against C. parvum infection and showed that it could be possible to design a passive immunization strategy against C. parvum based on the anti-P23 yolk antibody in animals and immunosuppressed humans.