Therapeutic effect of biosynthetic gold nanoparticles on multidrug-resistant Escherichia coli and Salmonella species isolated from ruminants

Research (Published online: 29-12-2021)

19. Therapeutic effect of biosynthetic gold nanoparticles on multidrug-resistant Escherichia coli and Salmonella species isolated from ruminants

Abeer M. Abdalhamed, Alaa A. Ghazy, Eman S. Ibrahim, Amany A. Arafa and Gamil S. G. Zeedan

Veterinary World, 14(12): 3200-3210

ABSTRACT

Background and Aim: Multidrug-resistant (MDR) pathogenic microorganisms have become a global problem in ruminants as a result of the intensive use of antibiotics, causing the development of resistance among gut microbiota. The antibiotic-resistant microorganisms can be transferred from diseased animals to humans. This study aimed to determine the prevalence of MDR Escherichia coli and Salmonella spp. isolated from cattle, buffaloes, sheep, and goats suffering from respiratory signs, diarrhea, and mastitis and to screen the antibiotic sensitivity of selected isolated bacteria. It also detected antibiotic-resistance genes by polymerase chain reaction (PCR), produced green gold nanoparticles (AuNPs) using plant extracts (Artemisia herba-alba and Morus alba), and evaluated the antimicrobial activities of these biosynthesized nanoparticles on selected pathogens (E. coli and Salmonella spp.).

Materials and Methods: MDR E. coli and Salmonella spp. were investigated using fecal samples (n=408), nasal swabs (n=358), and milk samples (n=227) of cattle, buffaloes, sheep, and goats with or without clinical signs, including respiratory manifestations, pneumonia, diarrhea, and mastitis, from different governorates in Egypt. E. coli and Salmonella spp. were isolated and identified on selective media, which were confirmed by biochemical reactions and PCR. Antimicrobial susceptibility testing against 10 commonly used antibiotics was performed using the Kirby-Bauer disk diffusion method. Antibiotic resistance genes blaTEM, blaSHV, blaOXA, and blaCTX-M were detected by PCR. The antibacterial effect of the biosynthesized AuNPs was evaluated by MIC and well diffusion assay. The biosynthesized AuNPs were also characterized by ultraviolet-visible spectrophotometry and transmission electron microscopy (TEM).

Results: Among all fecal samples, the prevalence of E. coli was 18.4% (183/993) and that of Salmonella spp. was 16.7% (66/408), as determined by cultural and molecular tests. All isolates of E. coli and Salmonella spp. were 100% resistant to ampicillin (AM) and amoxicillin and highly resistant to cefoxitin and AM-sulbactam. The total rate of resistance genes in E. coli was 61.2% (112/183), while that in Salmonella was 63.6% (42/66) for pathogens isolated from ruminants with respiratory manifestations, pneumonia, diarrhea, and mastitis. Among the resistance genes, blaTEM had the highest prevalence rate in E. coli (25.9%, 21/81) while blaSHV had the lowest (9.8%, 8/81) in fecal swabs. AuNPs were successfully synthesized using aqueous leaf extract of A. herba-alba and M. alba as bioreducing agents. TEM analysis showed particle size of 10-42 nm for A. herba-alba and M. alba AuNPs. The biosynthesized AuNPs showed antibacterial activity against MDR E. coli and Salmonella spp.

Conclusion: Rapid and accurate diagnostic methods are the cornerstone for effective treatment to reduce the risk of antimicrobial-resistant pathogenic microorganisms. This is particularly important for overcoming the increasing rate of MDR in ruminants with respiratory manifestations, pneumonia, diarrhea, and mastitis. This can be complemented by the development of AuNPs synthesized in an environmentally friendly manner AuNPs using natural plant extracts for the treatment of antibiotic-resistant microorganisms.

Keywords: Escherichia coli, gold nanoparticle, multidrug-resistant, ruminant, Salmonella species.

Validation of Brix refractometer to estimate immunoglobulin G concentration in goat colostrum

Research (Published online: 29-12-2021)

18. Validation of Brix refractometer to estimate immunoglobulin G concentration in goat colostrum

Chollada Buranakarl, Sumpun Thammacharoen, Morakot Nuntapaitoon, Sapon Semsirmboon and Kazuo Katoh

Veterinary World, 14(12): 3194-3199

ABSTRACT

Background and Aim: Immunoglobulin G (IgG) concentration is high in goat colostrum, particularly in the first few hours after parturition, and this is important for the kid's immunity and growth. IgG levels vary depending on several factors, including breed, disease status, colostrum management, handling, and collection time postpartum. A handheld optical refractometer, an affordable instrument that is simple to use in the field, is used widely in dairy farms to measure total solids. However, it can also be applied to estimate colostrum IgG content on the basis of comparison with standard measurement methods, usually radial immunodiffusion. Studies comparing %Brix values in relation to IgG concentration measured using enzyme-linked immunosorbent assay (ELISA) in goats are limited. The present study aimed to evaluate the use of a handheld optical Brix refractometer for the measurement of IgG concentration in goat colostrum, compare results with those using ELISA, and estimate the %Brix cutoff value equating to low-quality colostrum.

Materials and Methods: Colostrum samples were collected on day 0 from 21 goats (nine Black Bengal, six Saanen, and six of their crossbred offspring) and were frozen. Subsequently, they were analyzed for IgG concentration using a goat-specific ELISA test and Brix percentage using a handheld refractometer. The optimum %Brix cutoff value for the evaluation of colostrum quality was evaluated.

Results: The mean IgG concentration and %Brix in colostrum were 10.60±0.64 and 25.0±0.9 mg/mL, respectively. There was a significant (p<0.01) correlation between %Brix and IgG concentration. For an IgG concentration of 6.9 mg/dl, the cutoff value for %Brix was 18.5, equating to high specificity (100%) but low sensitivity (50%). A higher %Brix cutoff value of 21.5 showed high specificity (95%) and high sensitivity (100%).

Conclusion: A Brix refractometer can be used to estimate goat colostrum quality with a proposed %Brix cutoff value of <18.5%-21.5% for poor-quality colostrum.

Keywords: caprine, colostrum quality, optical Brix refractometer.

Effects of xylazine and adrenaline combinations: Preliminary clinical application for non-surgical protocols of nephrosplenic entrapment in horses

Research (Published online: 28-12-2021)

17. Effects of xylazine and adrenaline combinations: Preliminary clinical application for non-surgical protocols of nephrosplenic entrapment in horses

Worakij Cherdchutham, Patskit Sukhong, Kanchanog Sae-oueng, Nithisphat Supanwinijkul, Kittanai Wiangnak, Jirayut Srimuang, Tawanhathai Apichaimongkonkun, Sarocha Limratchapong and Soontaree Petchdee

Veterinary World, 14(12): 3188-3193

ABSTRACT

Background and Aim: The medical treatment of horses with nephrosplenic entrapment (NSE) of the large colon through administrating phenylephrine and rolling during general anesthesia was effective and less expensive than surgical treatment. However, the selection of drugs for non-surgical treatment of NSE is not a usual method for clinical practice. This study aimed to identify the effects of combined drugs on the cardiac and splenic response in horses and provide information on the NSE of the large colon for clinical application.

Materials and Methods: Six healthy Thai native crossbred horses were enrolled in this study. Horses received two protocols with a withdrawal period of 14 days: Group 1 received xylazine (0.5 mg/kg IV) and adrenaline (1 mcg/kg IV), and Group 2 received xylazine (0.5 mg/kg IV) and adrenaline (3 mcg/kg IV). Heart rate (HR), HR variability (HRV), heart dimensions, and the splenic response of six horses were measured before the sedation, 30 and 60 min later, and 65, 70, 75, 80, 90, and 100 min after adrenaline administration. Doppler was used to obtain systolic blood pressure.

Results: The HRV low-frequency and high-frequency power ratios decreased after using xylazine. Hypertension was observed after adrenaline administration. In this study, there were only minimal differences in the HR and respiratory rate between groups. However, overall cardiac and splenic parameters were statistically higher in Group 2.

Conclusion: This study suggested that xylazine and three micrograms of adrenaline preserved the cardiac autonomic activity balance and were safe to use non-surgical applicability in horses.

Keywords: adrenaline, autonomic nervous activity, horse, spleen contraction.

In silico molecular docking and in vitro analysis of ethanolic extract Ocimum sanctum Linn.: Inhibitory and apoptotic effects against non-small cell lung cancer

Research (Published online: 28-12-2021)

16. In silico molecular docking and in vitro analysis of ethanolic extract Ocimum sanctum Linn.: Inhibitory and apoptotic effects against non-small cell lung cancer

Ulayatul Kustiati, T. S. Dewi Ratih, N. Dwi Aris Agung, Dwi Liliek Kusindarta and Hevi Wihadmadyatami

Veterinary World, 14(12): 3175-3187

ABSTRACT

Background and Aim: Lung cancer, especially non-small cell lung cancer (NSCLC), has been identified as the leading cause of cancer deaths worldwide. The mortality rate from lung cancer has been estimated to be 18.4%. Until now, conventional treatments have not yielded optimal results, thus necessitating an investigation into the use of traditional herbal plants as potential candidates for its treatment. This study aimed to determine the inhibitory and apoptotic activity of the ethanolic extract from Ocimum sanctum Linn. (EEOS) by in silico molecular docking and through in vitro studies using NSCLC cells (A549 cell line).

Materials and Methods: Dried simplicia of Ocimum sanctum was converted into EEOS using the maceration method. Spectrophotometry was then employed to analyze the EEOS compound. The known main active compounds were further analyzed for inhibitory and apoptotic effects on gene signaling using in silico molecular docking involving the downloading of active compounds from PubChem and target proteins from the Protein Data Bank; the active compounds and proteins were then prepared using the Discovery Studio software v. 19.0.0 and the PyRX 0.8 program, interacted with the HEX 8.0.0 program, and visualized with the Discovery Studio Visualizer v. 19.0. Finally, an in vitro analysis was performed using an antiproliferative-cytotoxic test (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay in the NSCLC A549 cell line).

Results: The analysis revealed that the active compounds in the ethanolic extract were dominated by quercetin (flavonoids) (47.23% b/b) and eugenol (phenolic) (12.14% b/b). These active compounds interacted with the active sites (residual amino acids) of the αvβ3 integrin, α5β1 integrin, caspase-3, caspase-9, and vascular endothelial growth factor. Hydrogen bonds and Pi-cation and Pi-alkyl interactions were involved in the relationships between the active compounds and the active sites and thus may reveal an antioxidant property of the extract. Furthermore, in vitro analysis showed the inhibitory and antiproliferative effects of the EEOS against non-small cell cancer (A549).

Conclusion: Taken together, our data showed the ability of EEOS as an inhibitor and apoptotic agent for lung cancer; however, further research is needed to determine the exact mechanism of EEOS as an herbal medication.

Keywords: in vitro, lung cancer, molecular docking, Ocimum sanctum.

Analysis of culling reasons during the breeding cycle and lifetime performance: The strategy to remove crossbred Landrace and Large White sows under tropical climate

Research (Published online: 26-12-2021)

15. Analysis of culling reasons during the breeding cycle and lifetime performance: The strategy to remove crossbred Landrace and Large White sows under tropical climate

Ratchadaporn Boripun, Watcharapong Mitsuwan, Pawinee Kulnanan, Thotsapol Thomrongsuwannakij and Warangkana Kitpipit

Veterinary World, 14(12): 3170-3174

ABSTRACT

Background and Aim: Sow culling is an important practice in commercial swine production because it is directly associated with the economic efficiency of the breeding herd. This study was conducted to analyze the reasons for sow culling and quantify the factors affecting culling in crossbred Landrace and Large White sows under tropical climate.

Materials and Methods: A total of 4887 culled sows from one parent stock farm located in Ratchaburi province, Western Thailand, were examined in this study. Culling reasons were grouped into the following eight categories according to farm management: (1) Reproductive disorders, (2) old age, (3) low performance, (4) diseases, (5) lameness, (6) udder problems, (7) body condition, and (8) other illnesses. Logistic regression analysis was used to explore the relationship between culling sows and environmental factors. Effects of parity and season of culling were considered as fixed effects in a statistical model.

Results: Descriptive statistics indicated the following factors accounting for sow removals: Old age (34.93%, n=1707), reproductive disorders (29.32%, n=1433), low performance (12.62%, n=617), lameness (12.56%, n=614), diseases (4.8%, n=235), body condition (4.68%, n=229), udder problems (0.79%, n=39), and other illnesses (0.26%, n=13). Parity and season of culling were also found to have a significant effect on sow culling (p<0.05). The majority of culling sows in this population were of old age and high parity.

Conclusion: This study indicated that the purposeful culling of sows on this farm was within the targeted range. However, the incidence of reproductive disorders was too high and required further investigations.

Keywords: culling, parity, season, sow, tropical climate.

Impact of L-carnitine supplementation on the in vitro developmental competence and cryotolerance of buffalo embryos

Research (Published online: 26-12-2021)

14. Impact of L-carnitine supplementation on the in vitro developmental competence and cryotolerance of buffalo embryos

Mohamed M. M. El-Sokary, Al-Shimaa Al-H. H. El-Naby, Amal R. Abd El Hameed, Karima Gh. M. Mahmoud and T. H. Scholkamy

Veterinary World, 14(12): 3164-3169

ABSTRACT

Background and Aim: Despite many trials, buffalo embryos have poor cryosurvivability because of their high lipid content. L-carnitine was found to be a lipid-reducing agent when added to oocyte and embryo culture media. The study aimed to determine the most effective concentration of L-carnitine to improve the oocyte developmental competence and cryotolerance of buffalo embryos.

Materials and Methods: In vitro maturation and embryo culture media were supplemented with four concentrations of L-carnitine: 0 (control), 0.25, 0.5, and 1 mM. Good-quality embryos on 7 days were vitrified using mixtures of dimethyl sulfoxide and ethylene glycol at two concentrations (3.5 and 7 M).

Results: The result showed that the cleavage and morula rates were significantly (p<0.05) higher in the 0.5 mM group. Blastocyst rates were significantly (p<0.05) higher at both 0.5 and 1 mM. The rates of viable embryos directly after thawing were significantly (p<0.05) increased in the 0.5 mM group. No significant difference was found in embryos cultured for 24 h after warming among all the groups.

Conclusion: The addition of L-carnitine at a concentration of 0.5 mM to the culture media improves the oocyte developmental competence and cryotolerance of buffalo embryos directly after warming but not after 24 h of culture. Nevertheless, further studies must identify how L-carnitine exerts its beneficial micromechanisms.

Keywords: antioxidant, buffalo embryos, cryotolerance, L-carnitine.

Establishment of semen collection technique using electroejaculator and semen cryopreservation of Javan leopard (Panthera pardus melas Cuvier, 1809)

Research (Published online: 25-12-2021)

13. Establishment of semen collection technique using electroejaculator and semen cryopreservation of Javan leopard (Panthera pardus melas Cuvier, 1809)

Bongot Huaso Mulia, Ardyta Widianti, Jansen Manansang, Dedi Rahmat Setiadi, Vincentia Trisna Yoelinda, Taufiq Purna Nugraha, Ni Wayan Kurniani Karja and Raden Iis Arifiantini

Veterinary World, 14(12): 3156-3163

ABSTRACT

Background and Aim: The Javan leopard (Panthera pardus melas Cuvier, 1809) is a subspecies of Panthera pardus spp., spread across the African and Asian regions. Information on reproductive aspects is crucial for wild animals, including the Javan leopard. In this study, we aimed to develop electroejaculator (EE) techniques and evaluate cryopreservation success in Javan leopard semen.

Materials and Methods: The semen of four adult Javan leopards was collected once a week using EE. Placement of the EE probe in the rectum was performed after ultrasound imaging (ultrasonography) to determine the prostate body location. The semen obtained was then evaluated macroscopically and microscopically. Three Javan leopards were used for cryopreservation. The ejaculate was divided into two parts [i.e., one part diluted with AndroMed® (Minitüb, Tiefenbach, Germany) and the other part with Steridyl® (Minitüb)] at a 1:1 ratio immediately after collection and evaluation. The semen was then packed in a 0.25 mL MiniStraw® (Minitüb) then equilibrated at 4°C for 2 h. After equilibration, the straw was then frozen in liquid nitrogen vapor. Frozen semen was then stored in containers until further evaluation.

Results: The results showed that ejaculation response occurred at all levels of stimulation, while erections did not always occur. The fastest ejaculation and erection occurred at the fourth voltage. The macroscopic evaluation showed that the semen volume was 0.80±0.26 mL, cloudy white, pH 7.44±0.14, and with watery semen consistency. The microscopic evaluation showed that the sperm motility was 66.98±0.39%, with sperm viability of 75.6±1.79%. Sperm concentration was 62.17±46.95×106 mL–1 with a total concentration of 42.14±23.51×106 cells. Normal sperm morphology is only 40.72±6.26%.

Conclusion: This study concluded that the development of a semen collection technique using an EE preceded by imaging of the EE probe location using ultrasound was effective for the ejaculation of Javan leopards. The characteristics of the semen of the Javan leopard showed moderate semen volume, sperm motility, and viability. Javan leopard showed low sperm concentration and normal sperm morphology.

Keywords: electroejaculator, Javan leopard, semen characteristics, semen cryopreservation.