Acute-phase proteins and oxidative stress biomarkers in water buffalo calves subjected to transportation stress

The objective of the present study was to determine the effect of the transportation stress on water buffalo calves. A total of 50 buffalo calves (8?±?1 months old, 165?±?13 kg) were assigned to one of two equal groups; the first group represented clinically healthy non-transported calves (control non-transported group; n?=?25) whereas calves of the second group were subjected to transportation (transported group; n?=?25). Blood samples were collected from control non-transported calves and from transported calves immediately after unloading (post-transportation). The present findings indicated that the examined hematological and biochemical parameters were not significantly (P?≤?0.05) changed in transported calves when compared with the control non-transported group. Furthermore, serum concentration of the investigated acute-phase proteins (APP) namely, haptoglobin (0.37?±?0.01), serum amyloid A (75.43?±?2.11), and fibrinogen (7.51?±?0.25) were significantly (P?≤?0.05) higher in transported calves when compared with control calves (0.1?±?0.01 g/l, 23.9?±?0.56 mg/l, and 4.2?±?0.16 g/l), respectively. Lipid peroxidation represented as malonaldhyde (56.78?±?3.42) was higher significantly (P?≤?0.05), whereas antioxidant biomarkers in the form of nitric oxide (17.68?±?0.89) levels, and activities of superoxide dismutase (7.37?±?0.53) and reduced glutathione (5.25?±?0.95) were lower significantly (P?≤?0.05) in the serum of transported calves when all were compared with the control group (24.68?±?0.19 nmol/g Hb, 21.80?±?0.24 mmol/ml, 9.24?±?0.1 U/g Hb, and 7.23?±?0.21 mmol/l), respectively. Conclusively, the present study demonstrated that transportation were significantly enough to trigger changes in APP and oxidative stress biomarkers in buffalo calves.     

In vitro comparison of soybean lecithin-based extenders for cryopreservation of buffalo (Bubalus bubalis) semen

The objective of the present study was to compare the quality of frozen buffalo semen processed in tris-citric egg yolk extender with two different commercially available soybean lecithin-based extenders. Split pooled ejaculates, possessing more than 70 % visual sperm motility were divided in three aliquots and diluted in AndroMed®, Bioxcell®, or tris-citric egg yolk (TCE) extenders. Post-thaw motion characteristics, viability, plasma membrane integrity, acrosome morphology, and DNA integrity of buffalo sperm were studied after thawing and incubation for 4 h. Results demonstrate that total motility in frozen–thawed semen processed in TCE extender were similar when compared with AndroMed® or Bioxcell® extenders (P?>?0.05). Kinematic parameters, such as average path velocity and linearity index in the soybean lecithin-based extenders were comparatively superior to TCE extender. After 4 h of incubation, proportions of overall and progressive motility decreased in all extenders but were comparatively superior in soybean lecithin-based extenders than TCE extender (P?<?0.05). The proportion of post-thaw sperm viability, plasma membrane integrity and normal apical ridge remained similar in all extenders (P?>?0.05). The post-thaw sperm viability was comparatively superior in soybean lecithin-based extenders than TCE after incubation at 37 °C for 4 h (P?<?0.05). The type of extender had not been the significant effect on the percentage of spermatozoa with DNA damage after thawing and incubation for 4 h. In addition, our results suggest that soybean lecithin base extenders can be used as a substitute for tris-citric egg yolk extender in cryopreservation of buffalo semen.     

Development and Validation of a Simple Sensitive Enzyme Immunoassay (EIA) for GH Determination in Buffalo Plasma

Abstract

A simple and highly sensitive enzymeimmunoassay (EIA) for GH determination in buffalo plasma on microtitreplates using biotin-streptavidin amplification system and the second antibody coating was developed. Biotin was coupled to GH and used to bridge between streptavidin-peroxidase and immobilized antiserum in competitive assay. The EIA was carried out directly in 100 µL buffalo plasma. The GH standards ranging from 0.05 ng/well/100 µL to 12.8 ng/well/100 µL were prepared in hormone free plasma collected from an aged (>15 years) senile buffalo. The sensitivity of the EIA procedure was 50 pg/well GH, which corresponded to 0.50 ng/mL plasma; the 50% relative binding sensitivity was seen at 800 pg/well/100 µL. Plasma volumes for the EIA, viz., 25, 50, and 100 µL did not influence the shape of standard curve, even though a slight drop in the OD₄₅₀ was seen with higher plasma volumes. For the biological validation of the assay, 12 Murrah buffalo calves were used. Six of these were administered synthetic bovine growth hormone-releasing factor (10 µg/100 kg body weight, i.v., and the remaining six animals were administered sterile normal saline and kept as controls. Jugular blood samples were collected at ?60, ?45, ?30, ?15, ?10, ?5, 5, 10, 15, and 30 min and, thereafter, at an interval of 15 min using an indwelling jugular catheter, beginning 1h prior to GRF injection up to 8 h post treatment. In all animals, a peak of GH was recorded within 5 to 20 min of GRF administration, which confirms the biological validation of the EIA. To confirm homogeneity of buffalo GH with bovine GH, a parallelism test was conducted between the buffer standard curve of bovine GH and GH measured from serial dilution of buffalo plasma containing a high level of endogenous growth hormone.     

First molecular detection of porcine circovirus type 2 in bovids in China

For the worldwide pig industries, porcine circovirus type 2 (PCV2) is an economically important pathogen. At present, the prevalence of PCV2 is common in Chinese swine herds. However, there is little information on PCV2 prevalence in non-porcine animals in China, such as bovids. Therefore, the goal of this study is to obtain the firsthand prevalence data of PCV2 in bovids in China. Two hundred and eighty serum and muscle samples from dairy cows (n = 180), buffalo (n = 50), and yellow cattle (n = 50) were analyzed by PCR. The detection results show that PCV2 infections (16 %, 8/50) only exist in buffaloes. In addition, there are different PCV2 viral DNAs identified by differential PCR in the same buffalo sample. Nucleotide sequencing and phylogenetic analysis results based on partial ORF1 and ORF2 sequences suggest that PCV2 strains have genetic diversity in buffaloes and they are divided into three different genotypes (PCV2b, PCV2d, and PCV2e, respectively). Moreover, to our knowledge, the PCV2d and PCV2e genotypes have not been previously reported in bovids. Through this study, the firsthand data of PCV2 prevalence in bovids in China was documented.     

Genetic diversity of 3′ region of glycoprotein D gene of bovine herpesvirus 1 and 5

Bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5) are closely related alphaherpesviruses of cattle. While BoHV-1 is mainly associated with respiratory/genital disease and rarely associated with neurological disease, BoHV-5 is the primary agent of meningoencephalitis in cattle. The envelope glycoprotein D of alphaherpesviruses (BoHV-1/gD1 and BoHV-5/gD5) is involved in the early steps of virus infection and may influence virus tropism and neuropathogenesis. This study performed a sequence analysis of the 3′ region of gD gene (gD3′) of BoHV-1 isolates recovered from respiratory/genital disease (n = 6 and reference strain Cooper) or from neurological disease (n = 7); and from seven typical neurological BoHV-5 isolates. After PCR amplification, nucleotide (nt) sequencing, and aminoacid (aa) sequence prediction; gD3′ sequences were compared, identity levels were calculated, and selective pressure was analyzed. The phylogenetic reconstruction based on nt and aa sequences allowed for a clear differentiation of BoHV-1 (n = 14) and BoHV-5 (n = 7) clusters. The seven BoHV-1 isolates from neurological disease are grouped within the BoHV-1 branch. A consistent alignment of 346 nt revealed a high similarity within each viral species (gD1 = 98.3 % nt and aa; gD5 = 97.8 % nt and 85.8 % aa) and an expected lower similarity between gD1 and gD5 (73.7 and 64.1 %, nt and aa, respectively). The analysis of molecular evolution revealed an average negative selection at gD3′. Thus, the phylogeny and similarity levels allowed for differentiation of BoHV-1 and BoHV-5 species, but not further division in subspecies. Sequence analysis did not allow for the identification of genetic differences in gD3′ potentially associated with the respective clinical/pathological phenotypes, yet revealed a lower level of gD3′ conservation than previously reported.     

Erythroplastidcytosis in a Major Mitchell’s cockatoo (Lophochroa leadbeateri)

Avian erythrocytes are typically nucleated. However, a small proportion of anucleated erythrocytes, termed erythroplastids, may be encountered in the peripheral blood of birds. We document the presence of a large proportion (up to 47.6 %) of erythroplastids in the blood of a Major Mitchell’s cockatoo (Lophochroa leadbeateri) and consider the pathophysiology of this finding.     

Objective Assessment of Bradykinesia Estimated from the Wrist Extension in Older Adults and Patients with Parkinson’s Disease

Parkinson’s disease (PD) presents several motor signs, including tremor and bradykinesia. However, these signs can also be found in other motor disorders and in neurologically healthy older adults. The incidence of bradykinesia in PD is relatively high in all stages of the disorder, even when compared to tremor. Thus, this research proposes an objective assessment of bradykinesia in patients with PD (G _PD: 15 older adults with Parkinson’s disease, 65.3 ± 9.1 years) and older adults (G _HV: 12 healthy older adults, 60.1 ± 6.1 years). The severity of bradykinesia in the participants of G _PD was assessed using the Unified Parkinson’s Disease Rating Scale. Movement and muscular activity were detected by means of inertial (accelerometer, gyroscope, magnetometer) and electromyographic sensors while the participants performed wrist extension against gravity with the forearm on pronation. Mean and standard error of inertial and electromyographic signal parameters could discriminate PD patients from healthy older adults (p value <0.05). In discriminating patients with PD from healthy older adults, the mean sensitivity and specificity were respectively 86.67 and 83.33%. The discrimination between the groups, based on the objective evaluation of bradykinesia, may contribute to the accurate diagnosis of PD and to the monitoring of therapies to control parkinsonian bradykinesia, and opens the possibility for further comparative studies considering individuals suffering from other motor disorders.     

Erythrocyte-protective effect of sarpogrelate hydrochloride (Anplag®), a selective 5-HT2 receptor antagonist: an in vitro study

Sarpogrelate hydrochloride (Anplag^®; Mitsubishi Tanabe Pharma Corporation, Osaka, Japan) is a selective S₂-serotonergic receptor antagonist and is used as an antiplatelet agent. Sarpogrelate has been reported to increase erythrocyte deformability and suppress shear stress-induced hemolysis. In this study, we tested the protective effect of sarpogrelate on erythrocytes in an in vitro model. We compared the normalized index of hemolysis (NIH) between the group that was treated with sarpogrelate (n = 5) and the control group (n = 5) in a mock circulation under a shear force generated by a centrifugal pump. In the former group, 300 mg/day sarpogrelate was orally administered to a human volunteer for 2 days before blood collection. The NIH was significantly lower in the sarpogrelate group than it was in the control group (0.0012 ± 0.0009 vs. 0.0027 ± 0.0005 g/100 l, respectively; p = 0.016). Sarpogrelate suppressed shear stress-induced hemolysis in an in vitro model suggesting that sarpogrelate has a protective effect on erythrocytes under mechanical shear stress.     

Osmotic fragility of human erythrocytes in vitro using Vipera lebetina venom

The objective of this study was to investigate the in vitro effect of Vipera lebetina venom on human erythrocytes’ osmotic fragility with respect to different venom concentrations, time of incubation with the venom, initial volume of red blood cells, and presence of antivenom. In vitro osmotic fragility of human erythrocytes was determined using Dacie and Lewis method. Osmotic fragility is increased significantly after incubation with the venom at different concentrations. The maximal effect was detected at the venom concentration of 400 μg. Time of incubation with the venom was an important factor in hemolytic process while the initial cell volume has no significant effect. Incubation of envenomed erythrocytes for 120 min with antivenom did not reverse the changes in osmotic fragility induced by the venom. The study suggests that the venom could alter the susceptibility of erythrocytes to hemolysis when subjected to osmotic stress and the degree of hemolysis depends on venom concentration and time of intubation with the venom.     

The efficacy and safety of alphacypermethrin as a pour-on treatment for water buffalo (Bubalus bubalis) infested with Haematopinus tuberculatus (Phthiraptera: Haematopinidae)

The sucking louse Haematopinus tuberculatus (Burmeister 1839) is an ectoparasite of buffaloes, cattle, camels, and American bison. Alphacypermethrin (ACYP) is a pyrethroid insecticide commonly used to control arthropods of veterinary and public health interest. Therapeutics, such as antiparasitic compounds, is often administered to buffaloes based on dosage and intervals recommended for cattle because very few drugs have buffalo-specific label indications. A trial was conducted on 20 louse-infested buffaloes at a farm to assess the efficacy and safety of ACYP pour-on, at the manufacturer’s recommended dose for cattle, on buffaloes naturally infested by H. tuberculatus. Ten animals were assigned to ACYP-treated group (ACYP-group) and ten to untreated control group (C-group). On day 0, all ACYP-group buffaloes received alphacypermethrin pour-on. Louse counts were performed on days ?1, 7, 14, 21, 28, 35, 42, 49, and 56 at eight predilection sites on the skin of each buffalo. ACYP was completely effective (100 %) at day 7, highly effective (99.8 %) at day 14, and completely effective (100 %) from day 21 until the end of the study (day 56 post-treatment). During the trial, ACYP was well tolerated by all animals as there were no observed clinically adverse reactions. The results of this trial suggest that ACYP is an effective, safe, and user-friendly compound suitable for treatment of buffaloes with natural louse infestations.     

Possible relationship between Plasmodium falciparum ring-infected erythrocyte surface antigen (RESA) and host cell resistance to destruction by chemicals

Repeated incubation of Plasmodium falciparum culture in 0.015 % saponin solution for a total of 35 min destroys most of the uninfected cells, leaving only the ring-infected erythrocytes (RIEs). Parasites concentrated by this method can subsequently complete the asexual cycle and infect other erythrocytes. It is possible that resistance to saponin is mediated by one or more of the numerous parasite proteins present in the host erythrocyte membrane. We have found that schizonts are as susceptible as uninfected erythrocytes to saponin, indicating that the protective protein is parasite stage specific. Studies with cultured parasites have shown that ring-infected erythrocyte surface antigen (RESA) strengthens host erythrocyte membrane and protects against destruction. Therefore, we hypothesize that RESA could be involved in resistance to saponin. Here, we have carried out PCR test on RESA gene, using three different primers. One of them showed that P. falciparum isolates collected directly from infected humans and cultured only for a few days, or not at all, have amplicon sizes ranging from 372 to 510 bp. However, the amplicon size changed to 873 bp when in vitro growth was continued for one or more weeks. This genetic transformation precedes acquisition of the ability to confer saponin resistance to RIEs.     

Examining Bidirectional Pathways Between Exposure to Political Violence and Adolescent Adjustment in Northern Ireland

Research on social ecologies of political violence has been largely confined to cross-sectional tests of the impact of political violence on child adjustment, limiting perspectives on more nuanced causal pathways, including tests of reciprocal relations between exposure to political violence and child adjustment. Based on a four-wave longitudinal study, this research breaks new ground in assessing bidirectional relations between exposure to political violence in the form of experience with sectarian antisocial behavior and adolescents’ adjustment problems. The study included 999 mother–adolescent dyads selected from working-class neighborhoods in Belfast ranked in the bottom quartile in terms of social deprivation in Northern Ireland, with approximately 35–40 families recruited to participate from each neighborhood. Across the four annual waves of data, adolescents (52% female) were 12.18 (SD = 1.82), 13.24 (SD = 1.83), 13.62 (SD = 1.99), and 14.66 (SD = 1.96) years old. Cross-lagged path models were tested through R package lavaan with full information maximum likelihood. Reflecting a reciprocal pathway, adjustment problems related to higher reports of experience with sectarian antisocial behavior 1 year later. Boys’ experience with sectarian antisocial behavior related to greater adjustment problems 1 year later, but this reciprocal path did hold for the girls. These findings offer promising directions toward better modeling of dynamic relations between exposure to political violence and adolescent adjustment over time.     

Resting EEG Discrimination of Early Stage Alzheimer’s Disease from Normal Aging Using Inter-Channel Coherence Network Graphs

Amnestic mild cognitive impairment (MCI) is a degenerative neurological disorder at the early stage of Alzheimer’s disease (AD). This work is a pilot study aimed at developing a simple scalp-EEG-based method for screening and monitoring MCI and AD. Specifically, the use of graphical analysis of inter-channel coherence of resting EEG for the detection of MCI and AD at early stages is explored. Resting EEG records from 48 age-matched subjects (mean age 75.7 years)—15 normal controls (NC), 16 with early-stage MCI, and 17 with early-stage AD—are examined. Network graphs are constructed using pairwise inter-channel coherence measures for delta–theta, alpha, beta, and gamma band frequencies. Network features are computed and used in a support vector machine model to discriminate among the three groups. Leave-one-out cross-validation discrimination accuracies of 93.6% for MCI vs. NC (p < 0.0003), 93.8% for AD vs. NC (p < 0.0003), and 97.0% for MCI vs. AD (p < 0.0003) are achieved. These results suggest the potential for graphical analysis of resting EEG inter-channel coherence as an efficacious method for noninvasive screening for MCI and early AD.     

Sickling,cytomorphology and blood parameters in brown brocket deer (Mazama gouazoubira Fischer, 1814)

Many species of deer, especially North American and Eurasian deer, show a sickling phenomenon (a morphological change in the red blood cells) in in vitro studies, which was subsequently attributed to the existence of different forms of haemoglobin. The objective of this study was to investigate the sickling phenomenon and its cytomorphological basis and, simultaneously, to determine normal blood parameters in captive brown brocket deer (Mazama gouazoubira). Blood was obtained from 30 healthy individuals, 20 males and 10 females. Blood cell morphology was studied with both light and electron microscopic techniques. Determinations of haematological values were done by manual methods. We found the sickling phenomenon to be present in this species in the form of erythrocytes in crescent or holly leaf shape. The scanning electron microscope showed the polymerization of haemoglobin, and the electrophoresis revealed the existence of dimorphic haemoglobin. The number of erythrocytes was larger than reported previously (21.84?±?3.41?×?10⁶/μl), due to their extremely small size (~3 μm diameter, 18.25?±?3.15 fl volume). The leucocytes had a typical mammalian morphology. This is the first record of sickling from a Neotropical deer species.     

Description of the gamonts of a small species of<Emphasis Type="Italic"> Hepatozoon</Emphasis> sp. (Apicomplexa,Hepatozoidae) found in<Emphasis Type="Italic"> Crotalus durissus terrificus</Emphasis> (Serpentes,Viperidae)

A small species of the genus Hepatozoon found in a specimen of Crotalus durissus terrificus from the Botucatu region, São Paulo State, Brazil is described. The morphologic alterations induced in the snake’s erythrocytes by the presence of this parasite are described. Morphology and morphometric analyses were performed using the Qwin Lite 2.5 computerized image analysis system (Leica). The Hepatozoon possessed a small and short body (8.1±0.5 μm long and 3.8±0.4 μm wide), with round extremities. The cytoplasm varied from pale blue to basophilic and had no granulations. Its nucleus was large, occupied a large area of the cytoplasm, and was irregular in shape and not condensed. Despite its small size, this parasite induced important changes in the host cell. Total parasitemia observed was 56.6%.     

Protective effect of Thunbergia laurifolia extract on hemolysis during Plasmodium berghei infection

This study was aimed to investigate the efficacy of Thunbergia laurifolia leaf extract to protect hemolysis in mice infected with Plasmodium berghei. Aqueous leaf extract of T. laurifolia was freshly prepared, and total polyphenol was then measured using Folin-Ciocalteu reagent method. For in vivo test, ICR mice were given intraperitoneally with this extract (1,000 mg/kg) once a day for four consecutive days and subsequently inoculated with 1?×?10⁶ parasitized erythrocytes of P. berghei ANKA by intraperitoneal injection for 8 days. The results showed that hemolysis was inhibited as indicated by %hematocrit (%Hct) which was normal in infected mice treated with T. laurifolia extract. Untreated and pyrimethamine-treated controls showed decreasing %Hct. Moreover, no any toxic signs were observed in normal mice treated with this extract. We conclude that T. laurifolia leaf extract clearly protects hemolysis during P. berghei infection in mice.     

Adaptation to inflammatory rheumatic disease: Do illness representations predict patients’ physical functioning over time? A complex relationship

Patients with an inflammatory rheumatic disease (IRD), are often faced with significant limitations in physical functioning. Illness representations are a key-factor of their illness-related experience. Our aim was to examine (a) whether illness representations can predict or only reflect IRD patients’ physical functioning over time, and (b) the specific pathways through which representations and physical functioning at baseline are associated with representations and functioning at follow-up. Patients with rheumatoid arthritis (N = 54) or systemic lupus erythematosus (N = 58) participated in the two phases of the study, 1 year apart. According to the results, illness representations were rather predicted by physical functioning than the other way around. At the same time, illness representations at baseline and at follow-up seemed to form a chain that mediated the relation between physical functioning at baseline and 1 year later. These findings may help us better delineate the interplay between the ways patients understand their condition and adaptation to illness.     

Representativeness of two sampling procedures for an internet intervention targeting cancer-related distress: a comparison of convenience and registry samples

Internet interventions often rely on convenience sampling, yet convenience samples may differ in important ways from systematic recruitment approaches. The purpose of this study was to evaluate potential demographic, medical, and psychosocial differences between Internet-recruited and registry-recruited cancer survivors in an Internet-based intervention. Participants were recruited from a cancer registry (n = 80) and via broad Internet outreach efforts (n = 160). Participants completed a set of self-report questionnaires, and both samples were compared to a population-based sample of cancer survivors (n = 5,150). The Internet sample was younger, better educated, more likely to be female, had longer time since diagnosis, and had more advanced stage of disease (p’s < .001), and the registry-sample was over-represented by men and those with prostate or other cancer types (p’s < .001). The Internet sample also exhibited lower quality of life and social support and greater mood disturbance (p’s < .001). Understanding how convenience and systematic samples differ has important implications for external validity and potential for dissemination of Internet-based interventions.