Toxoplasma gondii is a eukaryotic parasite of the phylum Apicomplexa, which infects all warm-blood animals, including humans. In the present study, we examined sequence variation in dense granule 20 (GRA20) genes among T. gondii isolates collected from different hosts and geographical regions worldwide. The complete GRA20 genes were amplified from 16 T. gondii isolates using PCR, sequence were analyzed, and phylogenetic reconstruction was analyzed by maximum parsimony (MP) and maximum likelihood (ML) methods. The results showed that the complete GRA20 gene sequence was 1,586 bp in length among all the isolates used in this study, and the sequence variations in nucleotides were 0-7.9% among all strains. However, removing the type III strains (CTG, VEG), the sequence variations became very low, only 0-0.7%. These results indicated that the GRA20 sequence in type III was more divergence. Phylogenetic analysis of GRA20 sequences using MP and ML methods can differentiate 2 major clonal lineage types (type I and type III) into their respective clusters, indicating the GRA20 gene may represent a novel genetic marker for intraspecific phylogenetic analyses of T. gondii.
Animals ; Base Sequence ; Brazil ; China ; Deer ; *Genetic Variation ; Genotype ; Goats ; Humans ; Molecular Sequence Data ; Phylogeny ; Protozoan Proteins/*genetics/metabolism ; Sheep ; Swine ; Toxoplasma/classification/*genetics/isolation & purification/parasitology/physiology ; Toxoplasmosis/*parasitology ; Toxoplasmosis, Animal/*parasitology ; United States

Toxoplasma gondii is an opportunistic protozoan parasite that can infect almost all warm-blooded animals including humans with a worldwide distribution. Micronemes play an important role in invasion process of T. gondii, associated with the attachment, motility, and host cell recognition. In this research, sequence diversity in microneme protein 6 (MIC6) gene among 16 T. gondii isolates from different hosts and geographical regions and 1 reference strain was examined. The results showed that the sequence of all the examined T. gondii strains was 1,050 bp in length, and their A + T content was between 45.7% and 46.1%. Sequence analysis presented 33 nucleotide mutation positions (0-1.1%), resulting in 23 amino acid substitutions (0-2.3%) aligned with T. gondii RH strain. Moreover, T. gondii strains representing the 3 classical genotypes (Type I, II, and III) were separated into different clusters based on the locus of MIC6 using phylogenetic analyses by Bayesian inference (BI), maximum parsimony (MP), and maximum likelihood (ML), but T. gondii strains belonging to ToxoDB #9 were separated into different clusters. Our results suggested that MIC6 gene is not a suitable marker for T. gondii population genetic studies.
Amino Acid Sequence ; Animals ; Base Sequence ; Cats ; Cell Adhesion Molecules/chemistry/*genetics/metabolism ; Deer ; *Genetic Variation ; Genotype ; Goats ; Humans ; Molecular Sequence Data ; Phylogeny ; Protozoan Proteins/chemistry/*genetics/metabolism ; Sequence Alignment ; Sheep ; Swine ; Toxoplasma/classification/*genetics/isolation & purification/physiology ; Toxoplasmosis/*parasitology ; Toxoplasmosis, Animal/*parasitology

The compressive strength of the original bone tissue was tested, based on the raw human thigh bone, bovine bone, pig bone and goat bone. The four different bone-like apatites were prepared by calcining the raw bones at 800 degrees C for 8 hours to remove organic components. The comparison of composition and structure of bone-like apatite from different bone sources was carried out with a composition and structure test. The results indicated that the compressive strength of goat bone was similar to that of human thigh bone, reached (135.00 +/- 7.84) MPa; Infrared spectrum (IR), X-ray diffraction (XRD) analysis results showed that the bone-like apatite from goat bone was much closer to the structure and phase composition of bone-like apatite of human bones. Inductively Coupled Plasma (ICP) test results showed that the content of trace elements of bone-like apatite from goat bone was closer to that of apatite of human bone. Energy Dispersive Spectrometer (EDS) results showed that the Ca/P value of bone-like apatite from goat bone was also close to that of human bone, ranged to 1.73 +/- 0.033. Scanning electron microscopy (SEM) patterns indicated that the macrographs of the apatite from human bone and that of goat bone were much similar to each other. Considering all the results above, it could be concluded that the goat bone-like apatite is much similar to that of human bone. It can be used as a potential natural bioceramic material in terms of material properties.
Animals ; Apatites ; chemistry ; Biomechanical Phenomena ; Bone Substitutes ; chemistry ; Bone and Bones ; physiology ; Cattle ; Compressive Strength ; Goats ; Humans ; Microscopy, Electron, Scanning ; Swine ; X-Ray Diffraction

The reliability of a Korean black goat (Capra hircus coreanae) to detect estrus in Himalayan tahrs (Hemitragus jemlahicus) for an artificial breeding program was investigated. Estrus in six female Himalayan tahrs was synchronized using fluorogestone acetate (FGA) sponges. Thirteen days later, 200 IU of PMSG and 100 IU of hCG were injected before removing the sponges and simultaneously injecting 5 mg of PGF2alpha the next day. Penetration of the cervical canal and the thickness and location of red crayon marks were examined 40~43 h later. Two females treated with sponges containing 60 or 45 mg of FGA had estrogen levels of 8.7 and 11.1 pg/mL, respectively. No red marks were found on the backs of these two tahrs. The remaining females had higher levels of estradiol, and the red crayon marks were clearly shown. The cervical folds of these tahrs were readily penetrated and the insemination gun was smoothly inserted into the uterine body. In conclusion, a Korean domestic goat with its chest crayon-harnessed was successfully used to detect estrus of Himalayan tahrs. This technique might be utilized as a part of breeding programs for wild goats and avoid the need for a vasectomy of conspecific males.
Animals ; Breeding/methods ; Estradiol/blood ; Estrus/physiology ; Estrus Detection/*methods ; Estrus Synchronization/methods ; Female ; Goats/*physiology ; Male ; Progesterone/blood

Schmallenberg virus (SBV), a novel orthobunyavirus, was first isolated in 2011. SBV preferentially infects the central nervous system of cattle and sheep and causes fever, diarrhea, a drop in milk yields, congenital malformations and stillbirths. Until June 2014, more than 200 scientific publications regarding SBV have been published. Although more than 20 articles on SVB were published in China, most of these articles provided only a brief introduction of the disease without fully discussing the associated disease characteristics. As a new disease, it has been made a focus of the National Research Center for Exotic Animal Diseases at the China Animal Health and Epidemiology Center. In this review, in order to provide a reference for research into SBV in China, we have reviewed the state of current research progress on the etiology, diagnosis and epidemiology of SBV, and vaccine development.
Animals ; Bunyaviridae Infections ; diagnosis ; epidemiology ; veterinary ; virology ; Cattle ; China ; epidemiology ; Goats ; Host Specificity ; Orthobunyavirus ; classification ; genetics ; isolation & purification ; physiology ; Sheep

Congenital Neospora caninum infection was diagnosed in two Saanen goat kids from two distinct herds with a history of abortion and weak newborn goat kids in the Southern region of the State of Minas Gerais, Brazil. The first kid was weak at birth, had difficulty to rise and was unable to nurse. Gross lesions of porencephaly and hydrocephalus ex vacuo were seen. Multifocal necrosis, gliosis and non-supurative encephalitis were observed in the brain. Several parasitic cysts with a thick wall that reacted strongly only with polyclonal antiserum to Neospora caninum were seen in the cerebral cortex, brain stem and cerebellum. The second kid was born from a Neospora caninum seropositive mother that aborted in the last pregnancy. It was born without clinical signs. The diagnosis of neosporosis was based on antibody titer of 1:800 to N. caninum by indirect fluorescence antibody test obtained from blood collected before the goat kid ingested the colostrum and Neospora caninum DNA was detected by polymerase chain reaction and sequenced from placenta. This is the first report of neosporosis in goats in the southeast region of Brazil.
Animals ; Antibodies, Protozoan/immunology ; Brazil ; Coccidiosis/congenital/immunology/parasitology/*veterinary ; Female ; Goat Diseases/congenital/immunology/*parasitology ; Goats ; Molecular Sequence Data ; Neospora/genetics/immunology/*isolation & purification/physiology ; Pregnancy

BACKGROUNDCartilage injury has a very poor capacity for intrinsic regeneration. The cell-based treatment strategy for the cartilage repair using differentiated bone marrow mesenchymal stem cells (BMSCs) is, however, a promising approach to the chondral repair. This study was aimed to explore the chondrogenic potential of the goat BMSCs in the Transwell co-culture system and the poly-laetide-co-glycolide (PLGA) scaffolds.

METHODSThe BMSCs were isolated from the goat iliac crest while the chondrocytes were obtained from the goat's last costal cartilage. In the Transwell co-culture system, the BMSCs co-cultured with chondrocytes were designed as group A, whereas the goat's BMSCs induced with the chondrogenic medium were group B. Both groups A and B were the experimental groups, while group C that only contained BMSCs was the control group. In the PLGA scaffolds co-culture system, BMSCs were seeded into the PLGA scaffolds, which were suspended in the 24-well plate, and the control group was established by presence or absence of chondrocytes at the bottom of the 24-well plate. Toluidine blue staining, Alcian blue staining, collagen II immunofluoresence, collagen II immunochemical staining, collagen I, collagen II, COL2a Q-PCR and osteopontin Q-PCR were used to examine the chondrogenic conditions as well as the expressions of chondrogenic and osteogenic genes.

RESULTSCells isolated from the aspirates of the goat bone marrow proliferated rapidly and gained characteristics of stem cells in Passage 4. However, the differentiations of chondrocytes were not apparent in Passage 3. The results from Toluidine blue staining, collagen II immunofluoresence and PCR showed the transformation of BMSCs to chondrocytes in the Transwell co-culture system and PLGA scaffolds. Although the cartilage gene expressions were upgraded in both chondrogenesis group and co-culture system, the osteopontin gene expression, which represents osteogenic level, was also up-regulated.

CONCLUSIONSThe Transwell co-culture system and the PLGA scaffolds co-culture system can promote the chondrogenic differentiation of the goat's BMSCs, while up-regulated osteopontin gene expression in the Transwell co-culture system implies the osteogenic potential of BMSCs.

Animals ; Bone Marrow Cells ; physiology ; Cell Culture Techniques ; methods ; Chondrocytes ; physiology ; Chondrogenesis ; physiology ; Coculture Techniques ; Goats ; Mesenchymal Stromal Cells ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds

Oocytes retrieval, in vitro maturation (IVM) and fertilization (IVF) efficiency are inevitable steps towards in vitro production of embryos. In the present study, these parameters were investigated in the ovaries of prepubertal (n = 31) and pubertal (n = 61) black Bengal goats obtained from a slaughterhouse. Nuclear maturation was evaluated upon aspiration and following IVM in TCM-199 (Earle's salt with L-glutamine and sodium bicarbonate) for 27 h at 39degrees C under 5% CO2 in humidified air. The oocytes retrieval and efficiency (mean +/- SD) per prepubertal and pubertal goats were 5.2 +/- 0.6 and 6.8 +/- 0.6, and 77.3 +/- 0.1% and 80.5 +/- 0.6%, respectively. Anaphase I - telophase I stages differed significantly (7.3 +/- 0.8 vs. 2.6 +/- 0.2, p < 0.05) between the two groups of goats. After IVM, the percentages of metaphase II were significantly higher (66.3 vs. 60.3, p < 0.05) in pubertal goats than in their prepubertal counterparts. The percentages of normal in vitro fertilization (IVF) in Fert-Tyrode's albumin lactate pyruvate of pubertal goat oocytes did not differ between Percoll and swim-up sperm separation methods (36.7 +/- 0.9% vs. 32.7 +/- 1.3%, p > 0.05). Furthermore, sperm capacitation by heparin alone or in combination with ionomycin did not lead to a significant increase in the normal fertilization rate (34.8 +/- 1.7 vs. 32.2 +/- 1.5%, respectively) in the oocytes of pubertal goats. In conclusion, the ovaries of pubertal black Bengal goats obtained from the slaughterhouse could be used for in vitro embryo production. However, further optimization of the IVM and IVF techniques are necessary for satisfactory in vitro embryo production.
Animals ; Culture Techniques ; Embryonic Development/*physiology ; Female ; Fertilization in Vitro/*veterinary ; Goats/*embryology ; Male ; Oocyte Retrieval/veterinary ; Oocytes/*physiology ; Ovary/cytology ; Sexual Maturation ; Sperm Capacitation ; Tissue and Organ Harvesting/veterinary

OBJECTIVETo study biomechanical changes of newly formed bones 24 weeks after repairing large defects of long bones of goats using heterogeneous deproteinated bone (DPB) prepared by modified methods as an engineering scaffold.

METHODSAccording to a fully randomized design, 18 goats were evenly divided into three groups: normal bone control group (Group A), autologous bone group (Group B) and experimental group (Group C). Each goat in Groups B and C were subjected to the periosteum and bone defect at middle-lower part of the right tibia (20% of the whole tibia in length), followed by autologous bone or DPB plus autologous MSCs + rhBMP2 implantation, respectively and semi-ring slot fixation; while goats in Group A did not perform osteotomy. At 24 weeks after surgery, biomechanical tests were carried out on the tibias.

RESULTSAt 24 weeks after surgery, the results of anti-compression test on tibias in three groups were recorded by a functional recorder presented as linear pressure-deformation curve. The shapes of the curves and their change tendency were similar among three groups. The ultimate pressure values were 10.74 MPa+/-1.23 MPa, 10.11 MPa+/-1.35 MPa and 10.22 MPa+/-1.32 MPa and fracture compression rates were 26.82%+/-0.87%, 27.17%+/-0.75% and 28.22%+/-1.12% in Groups A, B and C, respectively. Comparisons of anti-compression ultimate pressures and fracture compression rates among three groups demonstrated no significant difference (P(AB) equal to 0.415, P(BC) equal to 0.494). Three-point anti-bend test on tibias was recorded as load-deformation curves, and the shapes of the curves and their change tendency were similar among three groups. The ultimate pressure values of the anti-bend test were 481.52 N+/-12.45 N, 478.34 N+/-14.68 N and 475.62 N+/-13.41 N and the fracture bend rates were 2.62 mm+/-0.12 mm, 2.61 mm+/-0.15 mm and 2.81 mm+/-0.13 mm in Groups A, B and C, respectively. There was no significant difference between groups (P(AB) equal to 0.7, P(BC) equal to 0.448). The ultimate anti-torsion torque values were 6.55 Nm+/-0.25 Nm, 6.34 Nm+/-0.18 Nm and 6.42 Nm+/-0.21 Nm and fracture torsion rates were 29.51 degree+/-1.64degree, 28.88 degree+/-1.46 degree and 28.81 degree+/-1.33 degree in Groups A, B and C, respectively. There was no significant difference between groups (P(AB) equal to 0.123, P(BC) equal to 0.346).

CONCLUSIONSThe biomechanical characteristics of newly formed bones from heterogeneous DPB for repairing large segmental long bone defect are comparable to those of normal bones and autologous bones. DPB has the potential for clinical usage as bone graft material.

Animals ; Biomechanical Phenomena ; Female ; Goats ; Male ; Proteins ; Tibia ; physiology ; surgery ; Tissue Engineering ; Torsion, Mechanical

A new kind of Cage made of poly-DL-lactic acid was designed and an in-vitro study was conducted to evaluate the biomechanical effect of PDLLA Cage on a goat cervical spine model. 27 goat cervical spines were divided into four groups randomly: intact group, PDLLA Cage group, titanium Cage group, and autologous tricortical iliac crest bone group. Different implants were implanted after complete discectomy (C3-4) was performed, then they were tested in flexion, extension, axial rotation, and lateral bending with a nondestructive stiffness method using a nonconstrained testing apparatus, and three-dimensional displacement was measured. The range of motion (ROM) and the mean stiffness values were calculated and compared between groups. The result showed that the ROM values between PDLLA Cage group and the titanium Cage group were not significantly different in extensional and rotational movement (P > 0.05), but they were lower than that of rest groups (P < 0.05). In flexional loading mode, the ROM values showed statistically significant difference between the four groups (P < 0.05). And in lateral bending loading mode, no significant difference was found between PDLLA Cage group and iliac crest bone group (P > 0.05), but significant difference was found among the other groups (P < 0.05). The stiffness of cervical spine was raised after Cage was implanted. In flexional and rotational loading mode, significant difference in stiffness was found between PDLLA Cage group and control group or iliac crest bone group (P < 0.05). So PDLLA Cage can provide enough primary stability for cervical intervertebral fusion.
Animals ; Biomechanical Phenomena ; Bone Transplantation ; Cervical Vertebrae ; physiology ; surgery ; Equipment Design ; Female ; Goats ; Joint Instability ; prevention & control ; Lactic Acid ; Orthopedic Fixation Devices ; Polyglycolic Acid ; Prostheses and Implants ; Spinal Fusion ; instrumentation