The Protective Role of Vitamin E against Oxidative Stress and Immunosuppression Induced by Non-Esterified Fatty Acids in Bovine Peripheral Blood Leukocytes.

High levels of non-esterified fatty acids (NEFAs) during the transition period lead to increased oxidative stress and immunosuppression in cows. Feeding them a vitamin-E-supplemented diet reduces reactive oxygen species (ROS) levels in the blood and diminishes immunosuppression in the transition period. However, whether the restoration of immune cell function occurs through the direct action of vitamin E in cells is still a topic that requires further discussion. Therefore, in this experiment, we aimed to investigate the effect of NEFAs on peripheral blood leukocytes (PBLs) and whether vitamin E mitigates the impact of NEFAs. We employed three groups: (1) blank, (2) NEFA only, and (3) pre-culturing with vitamin E before NEFA treatment (VENEFA). In peripheral blood mononuclear cells (PBMCs), there were no differences in vitamin E content among the three groups. However, in the vitamin E pre-treatment group, the vitamin E levels of polymorphonuclear neutrophils (PMNs) were significantly higher than those in the other two groups. NEFA levels increased malondialdehyde (MDA) levels in PBMCs, but pre-treatment with vitamin E reduced accumulation of MDA levels. Regarding the expression of proinflammatory genes, NEFAs increased the expression of interleukin-1ß in PBMCs and colony-stimulating factor 2 in PMNs. Vitamin E pre-treatment restored the increase in interleukin-1ß levels caused by NEFAs in PBMCs. None of the groups affected the phagocytosis of PMNs. Few studies have confirmed that NEFAs cause oxidative stress in bovine PBLs. In summary, this study found that NEFAs induce oxidative stress in PBLs and alter the expression of inflammation-related genes; meanwhile, vitamin E can reduce some of the effects caused by NEFAs. This result may suggest that vitamin E can assist bovine PBLs in resisting the immune suppression caused by an NEB during the transition period.

The First Nationwide Surveillance of Severe Fever with Thrombocytopenia Syndrome in Ruminants and Wildlife in Taiwan.

Since the first discovery of severe fever with thrombocytopenia syndrome virus (SFTSV) in China in 2009, SFTSV has rapidly spread through other Asian countries, including Japan, Korea, Vietnam and Pakistan, in chronological order. Taiwan reported its first discovery of SFTSV in sheep and humans in 2020. However, the prevalence of SFTSV in domestic and wildlife animals and the geographic distribution of the virus within the island remain unknown. A total of 1324 animal samples, including 803 domestic ruminants, 521 wildlife animals and 47 tick pools, were collected from March 2021 to December 2022 from 12 counties and one terrestrial island. The viral RNA was detected by a one-step real-time reverse transcription polymerase chain reaction (RT-PCR). Overall, 29.9% (240/803) of ruminants showed positive SFTSV RNA. Sheep had the highest viral RNA prevalence of 60% (30/50), followed by beef cattle at 28.4% (44/155), goats at 28.3% (47/166), and dairy cows at 27.5% (119/432). The bovine as a total of dairy cow and beef cattle was 27.8% (163/587). The viral RNA prevalence in ticks (predominantly Rhipicephalus microplus) was similar to those of ruminants at 27.7% (13/47), but wild animals exhibited a much lower prevalence at 1.3% (7/521). Geographically the distribution of positivity was quite even, being 33%, 29.1%, 27.5% and 37.5% for northern, central, southern and eastern Taiwan, respectively. Statistically, the positive rate of beef cattle in the central region (55.6%) and dairy cattle in the eastern region (40.6%) were significantly higher than the other regions; and the prevalence in Autumn (September-November) was significantly higher than in the other seasons (p < 0.001). The nationwide study herein revealed for the first time the wide distribution and high prevalence of SFTSV in both domestic animals and ticks in Taiwan. Considering the high mortality rate in humans, surveillance of other animal species, particularly those in close contact with humans, and instigation of protective measures for farmers, veterinarians, and especially older populations visiting or living near farms or rural areas should be prioritized.

The first discovery of severe fever with thrombocytopenia syndrome virus in Taiwan.

Severe fever with thrombocytopenia syndrome (SFTS), an emerging tick-borne zoonosis, has been rapidly spread in many Asian counties since 2010, which raises the great concern in East Asia. Nevertheless, the infection status of SFTS in Taiwan remains unclear. To investigate the existence of SFTSV in Taiwan, a total of 151 serum samples collected from 31 sheep, 63 bovine and 57 dogs were enrolled this study. Furthermore, 360 adult female Rhipicephalus microplus were also included. One-step RT-nested PCR and IgG ELISA were conducted to test SFTSV specific RNA and antibodies, respectively. The result provided the first evidence of the existence of SFTSV RNA and antibodies in ruminants and ticks in Taiwan.

Human chorionic gonadotropin (hCG)-induced ovulation occurs later but with equal occurrence in lactating dairy cows: comparing hCG and gonadotropin-releasing hormone protocols.

This study assessed the effects of two hormones, human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH), on ovulatory responses during different diestrous stages in lactating dairy cows. Estrous cycles of 21 cows were synchronized and were enrolled in stage 1 of the experiment. The cows were treated with a prostaglandin (PG) F2α analog either 9 to 10 days [mid-diestrus (MD) group] or 5.5 to 6.5 days [early-diestrus (ED) group] after synchronized ovulation (day 0 = first PGF2α administration). On day 2, the cows were administrated 250 µg GnRH or 3000 IU hCG. Ovulation was determined every 2 h from 24 to 36 h after GnRH or hCG administration, and then every 4 h up to 72 h until ovulation. Cows in stage 2 were administered these treatments in the reverse order. The results indicated that average ovulation times in cows treated with GnRH in the MD group (GnRH-MD group) and cows treated with GnRH in the ED group (GnRH-ED group) were 30.0 ± 1.0 h and 28.8 ± 0.4 h, respectively. However, ovulation times for cows treated with hCG in the MD group (hCG-MD group) and cows treated with hCG in the ED group (hCG-ED group) were 35.8 ± 4.6 h and 32.8 ± 2.2 h, respectively, and ovulation occurred significantly later in the hCG-treated groups than in the GnRH-treated groups. In summary, we found that hCG-induced ovulation occurred later than GnRH-induced ovulation regardless of different diestrous peroids; however, the two treatments did not differ in terms of percentage of ovulation.

Effect of GnRH on ovulatory response after luteolysis induced by two low doses of PGF2α in lactating dairy cows.

This study investigated the effect of gonadotropin-releasing hormone (GnRH) on ovulatory response after complete luteolysis induced by two low doses of prostaglandin (PG)F2α in lactating dairy cows. Cows (n = 18) ranging between 45 and 65 days in milk were recruited for synchronization by a modified Ovsynch-48 protocol (GnRH-7 days-375 µg PGF2α-1 day-250 µg of PGF2α-1 day-GnRH) over a total of 23 estrous cycles. Synchronized cows (n = 16) were randomly assigned to GnRH and Saline groups in stage 1 of the experiment after 9-10 days of ovulation in synchronization. On days 0 and 1 (day 0 = first PGF2α administration), cows were treated with 375 and 250 µg PGF2α, respectively. On day 2, cows in the GnRH and Saline groups were administered 250 µg GnRH or 2.5 mL of 0.9% saline, respectively. Serum progesterone (P4) levels were measured and changes in the corpus luteum (CL) were ultrasonically monitored daily from day 0-3 to assess complete luteolysis. Preovulatory follicle diameter and ovulatory response were evaluated by ultrasonography. In stage 2, cows were treated in a manner converse to that in stage 1. The synchronization rate was 69.6% (16/23). In stages 1 and 2, cows showed complete luteolysis with P4 concentration <1 ng/mL or remaining CL area <50%. Average ovulation time was 29.3 ± 0.5 h, which mostly occurred between 28 and 30 h after GnRH injection. However, all cows in the Saline group ovulated later than 36 h post-injection, with an average time of 52.7 ± 8.6 h. There was no difference in preovulatory follicle diameter between the two groups (16.8 ± 0.5 and 17.3 ± 0.5 mm for GnRH and saline groups, respectively). Although ovulation rate was not correlated with treatment, the rate within 48 h of GnRH injection (93.3%) tended to be higher compared with that in the Saline group (60.0%). Thus, GnRH administration increased ovulation rate following complete luteolysis induced by two low doses of PGF2α. These results indicate that this simple protocol for dairy cows is an effective alternative to timed artificial insemination programs in the field.

Effect of two low doses of prostaglandin F2α on luteolysis in dairy cows.

In this preliminary study, we determined the effect of a modified method involving the administration of two low doses of prostaglandin F2α (PGF2α) at an interval of 24 h on luteolysis in dairy cows, and compared it with the standard single-dose method. Twenty-six cows were assigned to three groups treated with two low doses (TLD group, n = 10), one standard dose (SD group, n = 10), and one low dose (OLD group, n = 6) on day 9 to 10 of the oestrous cycle (day 0 = the day of PGF2α administration). Their serum progesterone (P4) levels and corpus luteum (CL) sizes were measured daily from day 0 to 4 to assess CL regression. The results indicated that the proportion of complete luteolysis, indicating a P4 value ≤ 1 ng/mL on day 3, was higher in the TLD group (100.0%) than in the SD (60.0%) and OLD (66.7%) groups. Ultrasonically detected changes in the CL area correlated with the shifts in the P4 values in both the TLD and the SD groups. The remaining CL area was significantly smaller in the TLD group (17.8% ± 3.3%) than in the SD or OLD group on day 4. Thus, we concluded that the proportion of luteolysis in cows was increased with two low doses of PGF2α as compared to a single PGF2α dose, indicating the necessity of the second dose of PGF2α. However, further studies with larger sample sizes in the field are required.

The utilization of a commercial soil nucleic acid extraction kit and PCR for the detection of Clostridium tetanus and Clostridium chauvoei on farms after flooding in Taiwan.

Clostridial diseases are zoonoses and are classified as soil-borne diseases. Clostridium chauvoei and Clostridium tetani cause blackleg disease and tetanus, respectively. Since bacteria and spores are re-distributed by floods and then, subsequently, contaminate soils, pastures and water; the case numbers associated with clostridial diseases usually increase after floods. Because Taiwan is often affected by flood damage during the typhoon season, possible threats from these diseases are present. Thus, this study's aim is to apply a combination of a commercial nucleic acid extraction kit and PCR to assess the prevalence of Clostridia spp. in soil and to compare the positivity rates for farms before and after floods. The minimum amounts of Clostridium tetanus and Clostridium chauvoei that could be extracted from soils and detected by PCR were 10 and 50 colony forming units (cfu), respectively. In total, 76 samples were collected from the central and southern regions of Taiwan, which are the areas that are most frequently damaged by typhoons. Noteworthy, the positive rates for Clostridium tetanus and Clostridium chauvoei in Pingtung county after the severe floods caused by a typhoon increased significantly from 13.73 and 7.84% to 53.85 and 50.00%, respectively. This study for the first time provides the evidence from surveillance data that there are changes in the environmental distribution of Clostridium spp. after floods. This study indicates that screening for soil-related zoonotic pathogens is a potential strategy that may help to control these diseases.

Strategy for the treatment of puerperal metritis and improvement of reproductive efficiency in cows with retained placenta.

The objective of this study was to improve the reproductive efficiency of dairy cows with puerperal metritis (PM) subsequent to retained placenta (RP) using a two-step treatment strategy. A total of 188 postpartum cows, aged from 2 to 8 years, were utilised for 2 experiments. In Experiment 1, cows affected with RP/PM were randomly assigned to two treatment groups. Cows in Group A (n = 17) were treated with 600 mg of ceftiofur intramuscularly for 3 days followed by intrauterine lavage with 0.1% chlorhexidine and infusion with 0.5% povidoneiodine, while cows in Group B (n = 16) received two intrauterine infusions, first with 5 g of oxytetracycline and then with 0.5% povidone-iodine. Cows with normal postpartum findings were regarded as the healthy control group (n = 26). Ultrasonographic examination revealed that the ovarian activities including the appearance of a dominant follicle and days to first ovulation of the cows in Group A during the early postpartum period differed from those of Group B (P < 0.05), which coincided with the results of uterine swabbing for bacteriology. In Experiment 2, cows with normal postpartum findings were allocated to Group D (n = 78), which received an ovulation protocol (GnRH - 7 d PGF2α - 48 h hCG - 24 h AI) on day 50 ± 2 postpartum. Cows affected with PM were randomly divided into two groups, Group E (n = 25) combined the treatments applied in Groups A and D, while Group F (n = 26) repeated the treatment administered in Group E except for uterine lavage. The results indicated that the pregnancy rate within 150 days postpartum and the mean days open in Group E (76.0% and 106.3 ± 4.6 days, respectively) were significantly different from those in Group F (38.5% and 137.9 ± 10.9 days, respectively) (P < 0.05). This study suggests that reproductive efficiency could be improved by using the two-step treatment to regulate uterine involution and an early resumption of ovarian function in cows with RP/PM.

Suppurative meningitis in a 7-day-old Formosan sambar deer (Cervus unicolor swinhoei) caused by Escherichia coli.

This article describes the clinical and pathological features of an orphan 7-day-old, male Formosan sambar fawn that was hospitalized for treatment of weakness. The fawn had been deprived of colostrum and developed suppurative meningitis that was attributed to Escherichia coli.

Disseminated abscessation complicated with bone marrow abscess caused by Arcanobacterium pyogenes in a goat.

An 8-month-old, Nubian wether with a history of systemic illness was euthanatized for a pathological examination. At necropsy, the presence of disseminated abscessation and cellulitis in the limbs was noted. Other postmortem findings associated with the visceral disease in this animal included multiple abscess lesions, mainly in the lungs, kidneys, phalanxes and vertebrae. Histopathologically, lesions of arteriolitis were found as evidenced by bacterial embolisms in pulmonary and renal arteriola, indicating a bacteremia in the patient. Arcanobacterium pyogenes was consistently isolated from 8 lesions of abscessations, including the lesions of subcutaneous abscesses as well as bone marrow abscess in phalanxes and thoracic vertebrae. This is the first published report of disseminated arcanobacterial infection with bone marrow abscess of both the phalanxes and vertebrae in goat.

Association of increased serum acute-phase protein concentrations with reproductive performance in dairy cows with postpartum metritis.

BACKGROUND: It is known that quantification of bovine acute-phase proteins could routinely provide early diagnostic and prognostic information for monitoring herd health status. OBJECTIVE: The aim of this study was to test the hypothesis that haptoglobin (Hp) and serum amyloid A (SAA) have the potential for indicating uterine infection, mainly due to subclinical endometritis, which can seriously influence the reproductive performance in dairy cows. METHODS: A total of 264 serum samples were collected from 24 Holstein dairy cows at defined intervals from 1 week prepartum to 6 months postpartum. Clinically healthy animals (Group 1, n=6) were compared with 24 animals having acute puerperal metritis (Group 2, n=18) and with concentrations obtained from healthy heifers (baseline values, n=10). The concentrations of serum Hp and SAA were measured using a hemoglobin-binding assay and Western blot analysis, respectively. Reproductive performance was assessed as the number of days open and the conception rate at the end of the study. RESULTS: Upper cut-off values for Hp and SAA concentrations in heifers were 130.9 microg/L and 51.9 microg/mL, respectively. Hp concentrations in Group 2 were significantly higher than those in Group 1 at all time points (P<.001). Additionally, among the successfully pregnant animals, the number of days open was significantly higher in 6 cows with Hp >130.9 microg/mL (median=316 days) than in 11 cows with Hp < or =130.9 microg/mL (median=120 days) (P=.0024). CONCLUSIONS: Serum Hp and SAA concentrations can be used to recognize uterine infection in postpartum cows. The association between elevated Hp values and number of days open suggests Hp may also be a useful indicator of poor prognosis for reproductive performance.

Molecular characterization of enrofloxacin resistant Actinobacillus pleuropneumoniae isolates.

Enrofloxacin (ER) resistant Actinobacillus pleuropneumoniae strains emerged in Taiwan in 2002. The mechanism of ER resistance in A. pleuropneumoniae has not yet been reported. A total of 48 A. pleuropneumoniae isolates were obtained from the lungs of pigs with pleuropneumonia in Taiwan between September 2007 and April 2008. Twenty-nine isolates were found to be resistant to enrofloxacin. To understand the mechanisms of A. pleuropneumoniae's resistance to ER, enrofloxacin susceptibility of the isolates along with the mutations of the quinolone resistance-determining region (QRDR) of DNA gyrase and topoisomerase IV, qnr genes were analyzed. Enrofloxacin resistant isolates were found to carry at least one mutation in the QRDR of gyrA, leading to amino acid changes at codon 83 or 87. Efflux pump inhibitor (Phe-Arg-beta-naphthylamide) decreased enrofloxacin minimum inhibitory concentration 2-16-fold, suggesting participation of efflux in ER resistance. Plasmid mediated quinolone resistance genes qnr were not detected in these isolates. In conclusion, enrofloxacin resistance of A. pleuropneumoniae may be linked to multiple target gene mutations and active effluxs.

The reproductive performance of female Formosan sambar deer (Cervus unicolor swinhoei) in semi-domesticated herds.

This study documented the reproductive performance of 210 adult female Formosan sambar deer (FSD, Cervus unicolor swinhoei) from four semi-domesticated deer herds in Taiwan. An extensive analysis of 525 reproductive records from 2000 to 2008, including the conditions of estrus, gestation, and parturition was conducted. The mean+/-S.E.M. lengths of the estrous cycle, gestation, and fawning interval were 18.2+/-0.5d (n=56), 258.6+/-0.3d (n=160), and 369.9+/-2.3d (n=122), respectively. Hand breeding was performed between June and December (n=494), with the majority (93.1%) occurring between July and October (P<0.05). Fawning occurred from February to September (n=318), and most frequently (83.0%) between April and June (P<0.05). Pregnancy rate per mating in FSD hinds was 64.4%. There was a 1.3:1 male-to-female ratio at birth (P<0.05) among 320 fawns, and only two cases of twinning (0.63%). The postnatal mortality rate was 6.6% (21/320), and the mortality rate in fawns before weaning did not exceed 8% on any farm. Fecundity was enhanced by high pregnancy rates and high offspring survival rates. This study provides baseline information on reproductive performance of FSD, which should be valuable to veterinarians and deer industry personnel for management of FSD on farms in subtropical countries.

Phylogenetic analysis of canine parvovirus VP2 gene in Taiwan.

Canine parvovirus (CPV) is a non-enveloped virus with a single-stranded DNA genome and causes infectious enteritis in dog. In this study, 36 isolates of CPV infection were obtained in Taichung, Taiwan from 2003 to 2004. Using primers that can distinguish subtypes of CPV, we amplified part of viral VP2 gene by polymerase chain reaction (PCR) and the PCR product was sequenced; results demonstrated that two isolates could be classified as type 2a of CPV and the others were type 2b. The complete coding region of VP2 gene of type 2b was also sequenced, and phylogenetic analysis of these DNA sequences revealed that our Taichung isolate was close to the V-120, FPV-314, 97-008, Taiwan 9, LCPV-T1, and T4 isolates; however, because of the degeneracy of codons, the amino acid sequences of Taichung isolate was similar to that of the 97-008 isolate from Japan. It is known that two important amino acid residues (Asn-426 in type 2a and Asp-426 in type 2b; Ile-555 in type 2a and Val-555 in type 2b) are the determinants for the discrimination of type 2a or type 2b. After scrutinizing the complete VP2 gene of our Taichung isolate, we found the VP2 protein of the Taichung isolate did possess this molecular feature of type 2b virus. Previous studies reported that type 2a virus was the major type in Taiwan; our finding showed that CPV type 2b was the predominant type in the middle part of Taiwan. Moreover, a unique Ala-489 in VP2 of our Taichung isolate was found, contrasting to a Val-489 in the VP2 of other strains.

The effect of different extenders on post-thaw sperm survival, acrosomal integrity and longevity in cryopreserved semen of Formosan Sika deer and Formosan Sambar deer.

This study investigates the efficacy of five extenders in contributing to the outcome of semen cryopreservation in Formosan Sika and Sambar deer. Pooled semen (n=4) of six males of each breed was used. In Sika deer, semen collection rate was 96% (23/24) over all electro-ejaculations. Volume, sperm motility and sperm concentration of fresh ejaculates was 0.5+/-0.4 ml, 77+/-6% and 1471.3+/-940.0 x 10(6) ml(-1), respectively. Post-thaw motility in respective extender was A: 66+/-16%; B: 71+/-2%; C: 73+/-6%; D: 9+/-4% and E: 26+/-12% (mean+/-S.D.). In extender C (74+/-14%) more viable spermatozoa were preserved than in the others (A: 64+/-10%; B: 48+/-11%; D: 41+/-16%; E: 47+/-6%; P<0.05). Acrosomal integrity was not influenced by extender composition. Post-thaw motility did not decrease during a 4-h incubation period, irrespective of the extender used (P>0.05). In Sambar deer, semen collection rate was 88% (21/24) over all electro-ejaculations. Volume, sperm motility and sperm concentration of fresh ejaculates was 1.3+/-0.5 ml, 82+/-4% and 379.1+/-252.2 x 10(6) ml(-1), respectively. Post-thaw motility was in respective extenders A: 69+/-2%; B: 74+/-6%; C: 73+/-2%; D: 13+/-6% and E: 31+/-20%. Extenders B and C were superior (P>0.05) with respect to sperm motility. Similarly, post-thaw viability in extenders A (70+/-7%), B (76+/-7%) and C (79+/-2%) was higher than that D (25+/-19%) and E (29+/-17%) (P<0.01). Sperm acrosomal integrity was better preserved in extenders B (86+/-4%) and C (83+/-4%) than in extenders A (54+/-13%), D (39+/-22%) and E (46+/-22%) (P<0.05). Post-thaw sperm longevity in extender A reduced from 69 to 16% during incubation (P<0.05) whereas only a slight decrease was observed in the other extenders after 4 h. In conclusion these data show that egg-yolk-Tris-Tes-glycerol based extender C containing Equex STM paste is optimal for freezing semen of Formosan Sika deer while egg-yolk-Tris-citric acid-glycerol based extender B containing Equex and extender C are superior in semen cryopreservation to others for Formosan Sambar deer.

Serum haptoglobin concentration in cattle.

To obtain a basal concentration of serum Haptoglobin (Hp) in cattle in Taiwan, Hp concentrations were measured from serum samples collected from 10 healthy heifers, every week for one year. The values were also compared with those collected from 15 cows diagnosed with postpartum metritis. The heifers were successfully impregnated by artificial insemination six months after the tests. Hp concentrations were also measured in the serum collected from 11 other cows within 3 weeks after parturition. The Hp assay developed in this study gave a good correlation (r=0.893)with Western blotting. The Hp concentration of 454 serum samples from the 10 heifers had a mean value of 83.6 +/- 34.1 mg/l, and there was no significant difference among individual heifers. The basal value of Hp in heifers was calculated as less than 73.6 mg/l. No significant difference in Hp concentration was observed among the 10 heifers during cold and warm seasons (19.8 +/- 2.2 degrees C vs 27.3 +/- 1.4 degrees C), or before and after pregnancy. The mean serum Hp concentration from cows suffering from postpartum reproductive disorders was 1133.5 +/- 627.1 mg/l, which was significantly greater than the serum of healthy heifers and postpartum cows (104.6 +/- 61.0 mg/l) (P<0.05). These results demonstrate that Hp concentration may be a useful indicator for cows with postpartum reproductive disorders.