Goat Serum Research Articles

Differential Effects of Reproductive Hormones on the Developmental Potential of Oocytes in Hainan Black Lambs/Goats

Background: The study elucidated the effects of reproductive hormones on the developmental potential of oocytes in Hainan lamb/adult goats by analyzing the changes in the concentrations of reproductive hormones (FSH, LH, 17β-E2) in Hainan lamb/adult goat serum. Methods: The experiment used radioimmunoassay to detect follicle stimulating hormone (FSH), luteinising hormone (LH), oestradiol (17β-E2), concentrations in the serum of Hainan goats (3-month-old/18-month-old) during different oestrous cycles and to analyze the changing pattern of hormones. Result: The results showed that the number of follicles obtained in lambs was significantly higher than in adult goats (P less than 0.05), but the quality of follicles in lambs was poor and the number of available oocytes in them was lower than in adult goats. During the estrous cycle, adult black goats exhibit peak serum concentrations of FSH and LH on the 13th day, reaching 2.73 ng/mL and 5.69 ng/mL, respectively. In lamb goats, FSH reaches its peak concentration on the 15th day at 2.56 ng/mL, while LH peaks on the 13th day at 5.13 ng/mL. On the 15th day, both adult black goats and lamb goats experience the lowest serum concentrations of 17β-E2, at 4.42 ng/mL and 7.63 ng/mL, respectively. The serum concentrations of FSH and 17β-E2 in adult black goats are significantly higher than those in lamb goats (P less than 0.05) In summary, FSH and 17β-E2 can promote the maturation and development of oocytes in Hainan 3-month-old lamb goats, which can be injected with the corresponding concentration of hormones on the 15th day during the synchronized estrus in Hainan 3-month-old lamb goats, to improve the quality and quantity of oocytes, to better utilize the in vitro embryonic production performance of the young animals and to accelerate the acquisition of good livestock breeds.

A simulated sunlight-activated plastic mimic enzyme for dual-mode detection of D-penicillamine and serum iron

No reports disclosed that whether the plastic centrifuge tube with intrinsic photoenzyme activity can be used for the specific sensing of pharmaceuticals and biomarkers in biofluids so far. We herein report the plastic eppendorf (EP) tube as both reaction vessel and sunlight-activated plastic mimic enzyme for colorimetric/smartphone dual-mode detection of D-penicillamine (D-PA) and iron (Ⅲ) ions in goat and rabbit serums accordingly. The photooxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) combined with simulated sunlight-activated EP tube was selectively inhibited by adding D-PA, and the absorbance at 652 nm of oxidized TMB decreased. Good linearity between the absorbance variation (or R/(R + G + B)) and D-PA level ranging from 10 to 300 µM was obtained, and the limits of detection (LOD) were determined as 8.50 μM (colorimetric) and 6.92μM (smartphone). Furthermore, the polyvinyl alcohol hydrogel encapsulated TMB in EP tube as a smartphone biosensor had been triumphantly used to monitor D-PA level in goat serum. Interestingly, the adsorption band of photo-oxidized EP tube-TMB system inhibited by D-PA gradually increased as the enhancement of iron (Ⅲ) ions concentration. Based on the “on–off-on” effect, a sensitive and selective method was further used for selectively sensing iron (Ⅲ) in rabbit serum. Notably, the dual-mode sensing platforms exhibited outstanding detection performance, with LOD values as low as 0.50 µM and 0.65 µM respectively. In short, this work not only excavated a photoactivated-plastic mimics, but also developed a robust, low-cost, simple and sensitive biosensor for simultaneous detection of D-PA and Fe (Ⅲ) ions in biofluids. The current photoactivated sensor does not requires the introduction of exogenous catalyst except for simulated sunlight, plastic tube and chromogenic substrate.

Investigating Platelet Alpha Degranulation Defect in Patients with Gaucher Disease Using Immunofluorescence-Microscopy on Peripheral Blood Smears

Background: Gaucher disease (GD) is caused by mutations in the GBA1 gene, leading to a dysfunctional glucocerebrosidase (GCase) enzyme. This enzyme's inability to hydrolyze its substrate, glucocerebroside, results in its accumulation in lysosomes, primarily within macrophages. Bleeding tendency is a prominent feature in patients with GD, arising from thrombocytopenia, abnormal coagulation, and platelet dysfunction. In a previous study, we demonstrated that the main platelet dysfunction associated with the bleeding phenotype in GD was platelet α-degranulation. Study aim: To determine if the platelet α-degranulation defect is associated with α-granule deficiency, evaluated through peripheral blood smears by immunofluorescence microscopy. Methods: Blood smears were prepared from EDTA blood on SuperFrost slides, dried overnight at room temperature (RT), fixed in ice-cooled acetone at -30°C for 2 minutes, dried for 2-4 hours at RT, and stored at -30°C. On the day of testing, slides were warmed to RT, blocked with 10% normal goat serum for 30 minutes, and incubated with markers of α-granules, i.e., von Willebrand factor (rabbit anti-human von Willebrand factor) and P-selectin (mouse anti-human CD62P) for 1 hour in the dark. After washing, slides were incubated with secondary antibodies for 1 hour in the dark, washed, coated with glycerine, and stored at 2-4°C overnight. Fluorescence microscopy was performed using Alexa Fluor 488 and 568 for green and red emissions, respectively. All participants signed informed consent for biobanking. Results: We included 37 patients with GD at a median age of 30 years (range: 4-70 years), of whom 23 received GD-specific therapy. The median (range) platelet count was 184 x109/L (65-430 x109/L). Of these, 17 patients (45%) exhibited a low P-selectin response to thrombin receptor activating peptide (TRAP) and collagen, indicating an α-degranulation defect. In six of these patients, blood smears showed low α-granule markers compared to controls, four of whom were bleeders. The remaining 11 patients with low activity showed normal staining of α-granule markers, with only one being a bleeder. All patients with a normal response to P-selectin had normal staining of α-granule markers. The α-degranulation defect and α-granule deficiency were not related to age, platelet count, or GD-treatment status. Conclusions: Our data indicate that the platelet α-degranulation defect in patients with GD can be partially explained by α-granule deficiency. However, a low P-selectin response was also observed in the absence of α-granule deficiency, suggesting a defect in α-granule release. Further studies are needed to explore whether the accumulation of α-synuclein secondary to mutant GCase in patients with GD contributes to the abnormal α-granule release, as was shown in vitro. Understanding the cause of platelet dysfunction may help in managing the bleeding risks and in developing targeted treatments for GD.

Cattle, sheep, and goat humoral immune responses against SARS-CoV-2

Following the emergence of SARS-CoV-2 in late 2019, several species of domestic and wild animals have been found to be susceptible to SARS-CoV-2 infection through experimental inoculation and animal surveillance activities. Detection of SARS-CoV-2 specific antibodies in animals is an important surveillance tool since viral shedding in animals can only be detected for a short period of time. In this study, convenience serum samples were collected from 691 cattle, 698 sheep, and 707 goats from several regions in the United States, between 2019 to 2022. The samples were evaluated for the presence of SARS-CoV-2 specific antibodies using two commercial enzyme-linked immunosorbent assays (ELISA); one based on the inhibition of the SARS-CoV-2 receptor-binding domain (sVNT) and the other based on the nucleocapsid protein (N-ELISA) of SARS-CoV-2. Positive samples from the sVNT were additionally evaluated using a conventional virus neutralization test (VNT) employing the Wuhan-like SARS-CoV-2 USA/WA1/2020 isolate. Our results indicate that ∼1% (6/691) of cattle, ∼2% (13/698) of sheep, and ∼2.5% (18/707) of goat serum samples were positive when using the sVNT, whereas ∼4% of cattle (25/691) and sheep (27/698), and 2.5% (18/707) of goat serum samples tested positive with the N-ELISA. None of the sVNT positive cattle, sheep, or goat serum samples had detectable neutralizing antibody activity (<1:8) against the SARS-CoV-2 USA/WA1/2020 isolate by the VNT. Our results indicate low seropositivity in cattle, sheep, and goats in the U.S., indicating the importance to continue monitoring for SARS-CoV-2 prevalence in animal species that are in close contact with humans.

SEROPREVALENCE OF TOXOPLASMA GONDII IN GOATS FROM SOUTHWESTERN MISSISSIPPI, USA.

Parasitic infection with Toxoplasma gondii is prevalent in human and animal populations worldwide. Goat production for food and fiber has increased in popularity, and consumer demand for meat and dairy products has led to higher rates of human consumption in the United States. This trend has increased the importance of assessing the public health significance of these dietary commodities. The occurrence of T. gondii and its relevance to commercial goat production for the human food market in Mississippi has not been previously addressed. This study estimated the seroprevalence of T. gondii in breeding-age goats raised for human consumption from 4 counties in southwestern Mississippi. One hundred and forty-four goat serum samples were collected between August 2007 and April 2008 from Hinds (n = 55), Adams (n = 36), Yazoo (n = 36), and Copiah (n = 17) counties. The overall seroprevalence was 12.5% (18/144) and raises concern and justification for preventative measures including periodic surveillance of goat herds and production facilities to ensure a safe food commodity and food hygiene education for consumers.

274 Evaluation of recombinant Capripoxvirus antigens in the development of indirect enzyme linked immunosorbent assays

Abstract Capripoxvirus is a genus of DNA viruses that includes sheep pox virus, goat pox virus and lumpy skin disease virus (LSDV). These viruses are considered high consequence viruses due to the severe clinical disease leading to economic losses to production and impact on trade. The recent spread of LSDV in Europe and Asia has demonstrated the transboundary nature of LSDV. The vaccines used to control Capripoxvirus infections currently do not have the ability to differentiate infected from vaccinated animals (DIVA) and the enzyme linked immunosorbent assays (ELISA) available for detection of antibodies to Capripoxvirus infections are not ideal. Previous research has been done to evaluate different Capripoxvirus antigens using E. coli expressed antigens; however, there has been no prior evaluation of baculovirus expressed Capripoxvirus antigens for use in ELISA. The objective of the current research was to produce 6 recombinant Capripoxvirus antigens using a baculovirus expression vector system (BEVS) and to evaluate these antigens in ELISAs to test bovine, sheep, and goat serum. The sequences used to produce the recombinant antigens were based on a consensus obtained using reference genomes consisting of all three viruses within the genus. All six antigens were produced and purified in-house using BEVS followed by benchtop or FPLC purification (Figure 1). To date, the antigens have been fully validated for performance in indirect ELISA using sheep sera (Figure 2) and partially using cattle sera, with goat sera validation to follow. The ELISAs were developed and optimized using known reference sera as controls. The performance of the ELISAs developed was assessed in comparison with the only commercially available Capripoxvirus serological test. The results demonstrate that some of the antigens used for the ELISAs completely outperformed the commercial test. Several antigens were found to be useful for ELISAs to detect responses in sheep and cattle sera. The sensitivities and specificities of the different ELISAs for sheep sera ranged from 99.54% to 53.57% sensitivity and 99.54 to 94.95% specificity with likelihood ratios above 10 using receiver operating characteristic curve analysis, while the commercial kit had a 42.85% sensitivity using the same positive sheep serum panel evaluated. The development of these serological assays allows for the simultaneous evaluation of immunogenicity of Capripoxvirus antigens through the testing of serum from experimentally infected animals. This study demonstrates the proof of principle, that sensitive and specific ELISAs can be developed for Capripoxviruses.

SEROLOGICAL INCIDENCE OF CONTAGIOUS PLEUROPNEUMONIA OF GOATS IN TAJIKISTAN

The article presents data on serological monitoring of contagious pleuropneumonia of goats in goat farms in Tajikistan. As a result of selective serological studies using enzyme immunoassay (ELISA test), it was found that contagious pleuropneumonia of goats has a high incidence in farms of the Republic of Tajikistan and causes significant economic damage to goat farms.Enzyme immunoassay of 159 samples of blood serum of goats of local breed from 9 goat farms in 7 districts of the country seropositivity was found in 54 animal heads, which was 34%. The most widespread PDAs were noted in the goat farms of the Rudaki, A. Jami and Rasht districts, respectively 80.45 and 40%.

Genetic and environmental determinants of Immunoglobulin G in kid serum and adult colostrum of dairy goats

Immune competence is a key factor in the health and resilience of dairy ruminants. The aim of this study was to study immunoglobulin G concentration in serum shortly after birth and in colostrum of lactating animals as proxy of passive immune transfer and colostrum quality, respectively, and thus provide new health and resilience indicators for selection. Data from 424 female kids and 203 primiparous goats, reared on an INRAE experimental farm, were analyzed. IgG concentration was measured in kid serum and goat colostrum, respectively, using the radial immunodiffusion technique. Linear models were used to identify factors of variation in serum and colostrum IgG concentration, and health events were monitored for 5 weeks after birth to measure the impact of passive immune transfer on kids' health. We found a significant effect of serum IgG level on hindquarters cleanliness score (as a proxy of digestive troubles) and nasal and ocular discharge. As expected, the absorption of IgG in the days following birth is essential to ensure the passive transfer of immunity from the mother to the youngster, and provide immune protection. Genetic parameters were estimated for serum IgG concentration in kids and for colostrum characteristics in primiparous goats. Heritability of serum IgG in kids was not significantly different from zero. This low value, with large SE, may be due to the limited sample size and large environmental variability likely not accounted for. Especially, we could not adjust for the amount of IgG that was ingested by the kid, which can have a significant effect on serum IgG and possibly on heritability estimates. Heritability of colostrum traits were moderate, with estimates of 0.25 (±0.16), 0.35 (±0.16), and 0.25 (±0.16) for IgG concentration, IgG mass and colostrum volume, respectively. Genetic correlations with milk production showed high standard errors due to small numbers. However, estimates suggest a negative genetic association between production and IgG concentration in colostrum (rg = -0.44 ± 0.33), probably linked to a positive association with total colostrum volume. The significant heritabilities that were estimated for the first time demonstrate the existence of genetic control of colostrum quality in goats, and open up opportunities for selection in this species.

Molecular and serological investigations of Batai virus in cattle and goats in the border area of Yunnan, China (2021-2022).

Batai virus (BATV), a zoonotic pathogen transmitted by mosquitoes, infects vertebrates, including livestock, birds, and humans. Although BATV has been detected and isolated in mosquitoes in Yunnan Province, China, there have been no reports of livestock infection. Thus, we conducted a molecular and serological investigation of BATV in cattle and goat sera collected in spring and autumn from 2021 to 2022 in Honghe Prefecture, Yunnan Province, on the China-Vietnam border. We used indirect enzyme-linked immunosorbent assays and reverse transcription real-time PCR (RT-qPCR) to test 929 cattle and 973 goat serum samples. BATV antibodies were detected in 262/929 (28.2%) cattle and 263/973 (27.0%) goat serum samples. RT-qPCR did not detect BATV RNA. The positive rate of BATV serum antibodies in cattle and goats in Luxi County was higher compared with other areas, and it was also higher in autumn compared with spring, which may be related to climate, temperature, and mosquito density. Although our findings indicated the presence of BATV infection in livestock in the region, RT-qPCR did not detect BATV RNA. Therefore, BATV monitoring in cattle and goats should be heightened in autumn, and the scope of host monitoring should be expanded to clarify the hosts and vectors of BATV infection.

Aptamer and sodium alginate decorated graphene oxide composite material with ion responsiveness for Low-density lipoprotein trapping

The accumulation of excess Low-density lipoprotein (LDL) is strongly associated with the occurrence of heart failure, coronary artery disease and hypercholesterolaemia, and is a major factor in cardiovascular and cerebrovascular disease. Concerns about the ways to decrease LDL level have continuously arisen. In this study, an ionic stimulation-responsive composite (i.e., GO@Apt@SA) is prepared with modification of graphene oxide (GO) utilising LDL-aptamer (Apt) and sodium alginate (SA). The ion-responsive behaviour of GO@Apt@SA synergistically interacts with the specific recognition property of the aptamer, enabling adsorption of LDL with higher capacity and specificity. Under the optimal experimental conditions, the maximum adsorption capacity of GO@Apt@SA for LDL is 730.6 μg mg−1. Interestingly, the aptamer complementary chain could trigger the release of LDL with favourable elution efficiency, which competitively binds with LDL-specific aptamer to trigger LDL release. More importantly, GO@Apt@SA exhibits satisfactory adsorption performances for LDL in goat serum, meaning that the composite material and technology are available for the extraction of LDL from complex sample matrices.

Exposure of small ruminants to the Schmallenberg arbovirus in Germany from 2017 to 2018 – animal-specific and flock-management-related risk factors

The Schmallenberg virus (SBV), an emerging Orthobunyavirus of mainly ruminant hosts, caused a substantial epidemic in European ruminant populations between 2011 and 2013. The pathogen is transmitted by arthropod vectors (Culicoides spp.) and can cause reproductive disorders and severe malformations of the offspring or stillbirth. The present study aimed to assess SBV seroprevalence among German sheep and goats a few years after the first virus detection in the country (November 2011). In addition, an extensive risk factor analysis including host-specific and husbandry-related factors was implemented. Seroprevalence was determined by examining serum samples from 2759 sheep and 446 goats out of a total of 70 flocks across five German federal states. The samples were withdrawn in the period between 2017 and 2018. Using a commercial competitive ELISA, antibodies against SBV were detected in all 70 investigated flocks. A percentage of 60.1 % (1657/2759) of the sheep and 40.4 % (180/446) of the goat sera contained SBV antibodies. Generalized linear mixed modeling revealed significant effects of host species (sheep > goats), age (old > young) and sex (female > male) on SBV seroprevalence. For both species, also the farming purpose, and for goats, ectoparasite treatment and the presence of cattle on the farm played a role in terms of risk for SBV exposure. The observations from this study still emphasize a wide distribution of the pathogen in Germany. Nevertheless, the observed seroprevalence might not be sufficient to achieve effective herd immunity. Pinpointing risk factors identified susceptible populations for targeted vaccination programs to reduce potential animal losses caused by SBV.

Development and validation of enzyme-linked immunosorbent assay for anti-mouse pertussis immunoglobulin G using international reference anti-Bordetella pertussis mouse serum NIBSC 97/642.

In this study, an in-house enzyme-linked immunosorbent assay (ELISA) was developed and validated. The titer of ELISA was calculated using the reference line (RFL) method based on the standard curve drawn using the international reference anti-mouse serum NIBSC (National Institute for Biological Standards and Control) 97/642. In the development step, signal to noise was depicted to select the buffers that showed the most appropriate ratio. In the validation step, standard range, precision, dilution linearity, and specificity were confirmed, and RFL and parallel line (PLL) methods were compared in precision and dilution linearity. Coating concentration for plate was achieved at 0.1 µg/mL for pertussis toxin (PT), 0.15 µg/mL for filamentous hemagglutinin antigen (FHA), and 0.25 µg/mL for pertactin (PRN). The signal to noise ratio was 22.02 for PT, 14.93 for FHA, and 8.02 for PRN with 0.25% goat serum in phosphate-buffered saline (PBS) as a dilution buffer, and 2% skim milk in PBS as a blocking buffer. Based on the precision results, we assessed the lower limit of quantification by 1, 0.2, and 1.5 EU/mL concentration for PT, FHA, and PRN which met the ICH (International Council for Harmonization) M10 criteria of a 25% accuracy and total error of 40%. In specificity, homologous serum was spiked into heterologous serum and the accuracy met the criteria. There was no difference in the results between RFL and PLL calculations (p-value=0.3207 for PT, 0.7394 for FHA, 0.2109 for PRN). ELISA validated with RFL calculation method in this study is a relatively accurate assay for mouse humoral immunogenicity test.

Differences in the serum lipid profile of goats with varying autonomic nervous system tone

Goat farming is a promising branch of animal husbandry, as statistics show a rapid increase in global demand for goat milk and meat products. To meet market demand, new methodological approaches are needed to improve animal productivity while maintaining the quality of the raw materials. In this context, this study aims to determine the impact of autonomic nervous system tone on lipid metabolism in goats. The study involved Saanen goats, with the experimental groups formed based on the results of variation-pulsometric study and electrocardiography, followed by the determination of autonomic nervous system tone using the Baevsky method. The serum lipid profile of goats was studied using a LabLine-010 spectrophotometer. The following indicators were biochemically analysed in serum samples: content of triacylglycerols, cholesterol, high-density, low-density, and very-low-density lipoproteins, and the atherogenic index was calculated. It was established that the content of triacylglycerols was 23% lower (P&lt; 0.05) in sympathotonic animals and 41% lower (P&lt; 0.001) in vagotonic animals compared to normotonic animals. The level of lowdensity lipoproteins in the blood serum of sympathotonic animals decreased by 40% (P &lt; 0.05), and in vagotonic animals – by 76% (P&lt; 0.01), relative to normotonic animals. The content of highdensity lipoproteins in sympathotonic animals increased by 36% (P&lt;0.001), relative to normotonic animals. The level of cholesterol in the blood serum of goats differed by 16% (P &lt; 0.01) in sympathotonic animals and 15% (P&lt; 0.01) in vagotonic animals compared to normotonic animals. A significant influence of autonomic nervous system tone on the processes of maintaining lipid homeostasis in goats was determined. Taking into account the individual features of the influence of the autonomic nervous system on lipid metabolism when distributing animals will contribute to the effective adjustment of diets. After all, each animal has a different tone of the autonomic nervous system, which significantly affects the features of substance transformation in its body

Immunisation of the somatostatin gene alters hypothalamic-pituitary-liver gene expressions and enhances growth in Dazu black goats.

Somatostatin (SS) plays important regulatory roles in animal growth and reproduction by affecting the synthesis and secretion of growth hormone (GH). However, the mechanism by which SS regulates growth and development in goats is still unclear. In this study, we randomly selected eight 7-month-old Dazu black goats (DBGs) of similar body weight and equally assigned four bucks as the immunised and negative control groups. The immunised group received the Salmonella typhi attenuated vaccine X9241 (ptCS/2SS-asd) orally, whilst the negative control group received the empty vector vaccine X9241 (pVAX-asd) orally. The SS concentration in the serum of goats in the immunised group was significantly lower than that in the negative control group, and the daily gain was significantly higher (p<0.05). SS-14 DNA vaccine immunisation resulted in significantly higher concentrations of growth-related hormones such as GH-releasing hormone and insulin growth factor 1 (IGF-1) in the serum of goats (p<0.05). RNA-seq analysis of hypothalamus of oral SS-14 DNA vaccine and negative control DBGs identified 31 differentially expressed genes (DEGs). Pituitary gland identified 164 DEGs. A total of 246 DEGs were detected in the liver by RNA-seq. Gene ontology of DEGs was enriched in mitochondrial envelope, extracellular region, receptor binding and cell proliferation. The biological metabolic pathways associated with DEGs were explored by Kyoto encyclopedia of genes and genomes analysis. DEGs were associated with metabolic pathways, oxidative phosphorylation, vitamin digestion and absorption and galactose metabolism. These candidate genes (e.g. DGKK, CYTB, DUSP1, and LRAT) may provide references for exploring the molecular mechanisms by which SS promotes growth and development. Overall, these results demonstrated that the SS DNA vaccine enhanced the growth of DBGs by altering growth-related hormone concentrations and regulating the expression of growth-related genes in the hypothalamic-pituitary-liver axis.

Detection of Ovarian Cancer Biomarker Lysophosphatidic Acid Using a Label-Free Electrochemical Biosensor

Electrochemical biosensors are valued for their sensitivity and selectivity in detecting biological molecules. Having the advantage of generating signals that can be directly or indirectly proportional to the concentration of the target analyte, these biosensors can achieve specificity by utilizing a specific biorecognition surface designed to recognize the target molecule. Electrochemical biosensors have garnered substantial attention, as they can be used to fabricate compact, cost-effective devices, making them promising candidates for point-of-care testing (POCT) devices. This study introduces a label-free electrochemical biosensor employing a gold screen-printed electrode (SPE) to detect lysophosphatidic acid (LPA), a potential early ovarian cancer biomarker. We employed the gelsolin–actin system, previously introduced by our group, in combination with fluorescence spectrometry, as a biorecognition element to detect LPA. By immobilizing a gelsolin–actin complex on an SPE, we were able to quantify changes in current intensity using cyclic voltammetry and differential pulse voltammetry, which was directly proportional to the LPA concentration in the solution. Our results demonstrate the high sensitivity of the developed biosensor for detecting LPA in goat serum, with a limit of detection (LOD) and a limit of quantification (LOQ) of 0.9 µM and 2.76 µM, respectively, highlighting its potential as a promising tool for early-stage diagnosis of ovarian cancer.

Changes in ovarian tissue structure and distribution of oestrogen receptors in Huanghuai goats at different ages.

To observe developmental changes in the ovarian tissue structure and distribution characteristics of oestrogen receptors (ERs) in the ovaries of Huanghuai goats at different ages, we selected healthy Huanghuai goats ewes and divided them into five groups (i.e. 3-, 30-, 60-, 90- and 120-day-old groups), with 10 animals in each group. The serum was separated after blood collection through the jugular vein, and the contents of oestrogen (E) and progesterone (P) in the serum of Huanghuai goats at each age were determined. Three goats were randomly selected from each group and sacrificed after anaesthesia, and the ovarian tissue was quickly obtained and placed in 4% paraformaldehyde fixative to prepare the tissue sections. Using HE, oestrogen receptors were immunohistochemically stained and observed. These results showed many primordial follicles and occasional secondary follicles in the ovaries of 3-day-old Huanghuai goats. Ovarian reticular structures were observed in 30-day-old ovarian medulla, with occasional near-mature growing follicles. Mature follicles and corpus luteum were occasionally detected in 60-day-old ovarian cortex. The 90-120-day-old ovarian cortices contained growing and mature follicles, and the number of mature follicles and corpora lutea increased, implying a significant luteal involution period. The E and P contents in the 120-day-old group were significantly higher than those in the 3-, 30-, 60- and 90-day-old groups. The levels of ERα and ERβ in the 3- and 30-day-old groups were mainly distributed in the granulosa cells of ovarian reproductive epithelial cells, primordial follicles, atretic follicles, and primary and secondary follicles. The ERα and ERβ levels of the 60-, 90- and 120-day-old groups were also distributed in the granulosa cells and luteal cells of mature follicles, especially in the 120-day-old endometrial cells of mature follicles, where ERβ was distributed significantly. The overall expression of ERβ in the ovary was higher than that of ERα. The results of this study provide basic data on the ovarian development and the specific expression of ERs and PRs in the ovaries of Huanghuai white goats, which play an important role in ovarian development and precocity.

Seroprevalence and risk factors of Toxoplasma gondii in goats (Capra hircus) in the state of Espírito Santo, Southeast Brazil

Toxoplasma gondii is described as a potential cause of abortion in goats and as a threat to public health. To estimate the prevalence of goats infected by T. gondii, in different cities in the Espírito Santo State, and to identify possible risk factors for infection a serological study was conducted. A total of 146 goat serum samples from the cities of Cariacica, Serra and Vila Velha were analyzed. The presence of IgG Class Immunoglobulins was serologically evaluated by Immunofluorescence antibody test (IFAT) and by Enzyme-linked Immunosorbent Assay (ELISA). The seroprevalence of anti-T. gondii was 46.6% (68/146) in both techniques and the same samples got the same results in both techniques. Among the analyzed sera, 70.6% (48/68) exhibited high-avidity IgG antibodies, and 29.4% (20/68) exhibited low-avidity IgG antibodies, suggesting that the infection was chronic in the infected animals. Female sex, age group over two years old, water from the public supply system, storage of food and supplies in an open and unprotected place, and the presence of a domestic cat on the property were identified as risk factors for T. gondii infection in goats. The state of Espirito Santo has a high frequency of infected goats, and this is the first research on caprine toxoplasmosis seroepidemiology in that region.

Facile fabricate sandwich-structured molecularly imprinted dopamine polymer for simultaneously specific capture of Low-density lipoprotein and eliminate “bad cholesterol”

A simplified approach for preparation of sandwich type molecularly imprinted polymers (PPDA-MIPs) is proposed for simultaneously identify Low-density lipoprotein (LDL) and dispose “bad cholesterol”. Porous polydopamine nanosphere (PPDA) is applied as a matrix for immobilization of LDL, and the imprinted layer is formed by dopamine acting as a functional monomer. Since imprinted cavities exhibit shape memory effects in terms of recognizing selectivity, the PPDA-MIPs exhibit excellent selectivity toward LDL and a substantial binding capacity of 550.3 μg mg−1. Meanwhile, six adsorption/desorption cycles later, the adsorption efficiency of 83.09 % is still achieved, indicating the adequate stability and reusability of PPDA-MIPs. Additionally, over 80 % of cholesterol is recovered, indicating the completeness of “bad cholesterol” removal in LDL. Lastly, as demonstrated by gel electrophoresis, PPDA-MIPs performed satisfactory behavior for the removal of LDL from the goat serum sample.

The Development of Toxoplasma gondii Recombinant Trivalent Chimeric Proteins as an Alternative to Toxoplasma Lysate Antigen (TLA) in Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of Immunoglobulin G (IgG) in Small Ruminants.

This study presents an evaluation of seventeen newly produced recombinant trivalent chimeric proteins (containing the same immunodominant fragment of SAG1 and SAG2 of Toxoplasma gondii antigens, and an additional immunodominant fragment of one of the parasite antigens, such as AMA1, GRA1, GRA2, GRA5, GRA6, GRA7, GRA9, LDH2, MAG1, MIC1, MIC3, P35, and ROP1) as a potential alternative to the whole-cell tachyzoite lysate (TLA) used in the detection of infection in small ruminants. These recombinant proteins, obtained by genetic engineering and molecular biology methods, were tested for their reactivity with specific anti-Toxoplasma IgG antibodies contained in serum samples of small ruminants (192 samples of sheep serum and 95 samples of goat serum) using an enzyme-linked immunosorbent assay (ELISA). The reactivity of six recombinant trivalent chimeric proteins (SAG1-SAG2-GRA5, SAG1-SAG2-GRA9, SAG1-SAG2-MIC1, SAG1-SAG2-MIC3, SAG1-SAG2-P35, and SAG1-SAG2-ROP1) with IgG antibodies generated during T. gondii invasion was comparable to the sensitivity of TLA-based IgG ELISA (100%). The obtained results show a strong correlation with the results obtained for TLA. This suggests that these protein preparations may be a potential alternative to TLA used in commercial tests and could be used to develop a cheaper test for the detection of parasite infection in small ruminants.

Spatially confined CuFe2O4 nanosphere in N/O-codoped porous carbon mimetics for triple-mode sensing of antibiotics and visual detection of neurotransmitters in biofluids

BackgroundCarbon-based nanozymes have recently received enormous concern, however, there is still a huge challenge for inexpensive and large-scale synthesis of magnetic carbon-based “Two-in-One” mimics with both peroxidase (POD)-like and laccase-like activities, especially their potential applications in multi-mode sensing of antibiotics and neurotransmitters in biofluids. Although some progresses have been made in this field, the feasibility of biomass-derived carbon materials with both POD-like and laccase-like activities by polyatomic doping strategy is still unclear. In addition, multi-mode sensing platform can provide a more reliable result because of the self-validation, self-correction and mutual agreement. Nevertheless, the use of magnetic carbon-based nanozyme sensors for the multi-mode detection of antibiotics and neurotransmitters have not been investigated. ResultsWe herein report a shrimp shell-derived N, O-codoped porous carbon confined magnetic CuFe2O4 nanosphere with outstanding laccase-like and POD-like activities for triple-mode sensing of antibiotic d-penicillamine (D-PA) and chloramphenicol (CPL), as well as colorimetric detection of neurotransmitters in biofluids. The magnetic CuFe2O4/N, O-codoped porous carbon (MCNPC) armored mimetics was successfully fabricated using a combined in-situ coordination and high-temperature crystallization method. The synthesized MCNPC composite with superior POD-like activity can be used for colorimetric/temperature/smartphone-based triple-mode detection of D-PA and CPL in goat serum. Importantly, the MCNPC nanozyme can also be used for colorimetric analysis of dopamine and epinephrine in human urine. SignificanceThis work not only offered a novel strategy to large-scale, cheap synthesize magnetic carbon-based “Two-in-One” armored mimetics, but also established the highly sensitive and selective platforms for triple-mode monitoring D-PA and CPL, as well as colorimetric analysis of neurotransmitters in biofluids without any tanglesome sample pretreatment.