Animal Samples Research Articles

Is green metallic sheen in emb agar accurate to assist the Escherichia coli identification?

Escherichia coli is an essential bacteria as a human and animal pathogen, although this species can also be disseminated in the environment. The clinical laboratories use EMB agar (Eosin Methylene Blue) to identify E. coli from the observation of metallic green sheen. However, other bacterial species may show sheen when sown on EMB agar, or in some cases, the pH sample influences the presence of this sheen by E. coli. In this study, MALDI-TOF was used as the gold standard to detect E. coli from human, animal, and environmental samples by analyzing the presence of green glow on EMB agar. The positive predictive values (PPV) ​​were higher (above 85%) in human, animal, and environmental samples studied, and the negative predictive value (NPV) was variable between the samples. Specificity and sensitivity were also variables suggesting the influence of the pH of the samples in the presence of metallic green brightness. This work demonstrated that it is necessary to implement biochemical tests or, when possible, molecular assays to confirm E. coli since only the observation of the production of metallic green glow in EMB can generate false negative results in the identification of E. coli. Keywords: Biochemical tests; Escherichia coli; Laboratory Analysis; MALDI-TOF.

Diversity and Potential Cross-Species Transmission of Rotavirus A in Wild Animals in Yunnan, China

Rotavirus A (RVA) is the primary enteric pathogen of humans and many other species. However, RVA interspecies transmission remains poorly understood. In this study, we conducted a comprehensive screening and genotyping analysis of RVA in 1706 wild animal samples collected from various regions within Yunnan Province, China. A total of 24 samples, originating from wild boars, rodents, bats, and birds tested positive for RVA. Next generation sequencing and phylogenetic analyses revealed a high degree of genetic diversity and reassortment, particularly for VP4 and VP7. Strains isolated from wild boars and rodents exhibited gene segments with high similarity to those found in humans and other mammalian RVA strains, indicating that RVA may undergo interspecies transmission and reassortment, resulting in novel strains with potential risks for human infection. This study provides critical data for understanding the transmission mechanisms and the RVA host range, and highlights the pivotal role of wildlife in viral evolution and dissemination. These findings have significant implications for public health policies and emphasize the need for enhanced surveillance to prevent interspecies RVA transmission.

Generating Virtual Bone Scans for the Purpose of Investigating the Effects of Cortical Microstructure.

Evaluating the contribution of microstructure to overall bone strength is tricky since it is difficult to control changes to pore structure in human or animal samples. We developed an open-source program that can generate three-dimensional models of micron-scale cortical bone. These models can be highly customized with a wide array of variable input parameters to allow for generation of samples with high similarity to CT scans of cortical bone or with specific geometric features. The program can generate samples with specific desired porosities (p < 0.001) and minor deviations in pore diameter from human samples: 2.29% (±4.61) using literature values, and 0.87% (±1.39) with optimized values.

Emerging biofilm formation and disinfectant susceptibility of ESBL-producing Klebsiella pneumoniae

Klebsiella pneumoniae is an opportunistic pathogen responsible for various infections in humans and animals. It is known for its resistance to multiple antibiotics, particularly through the production of Extended-Spectrum Beta-Lactamases (ESBLs), and its ability to form biofilms that further complicate treatment. This study aimed to isolate and identify K. pneumoniae from animal and environmental samples and assess commercial disinfectants’ effectiveness against K. pneumoniae isolates exhibiting ESBL-mediated resistance and biofilm-forming ability in poultry and equine farms in Giza Governorate, Egypt. A total of 320 samples, including nasal swabs from equine (n = 60) and broiler chickens (n = 90), environmental samples (n = 140), and human hand swabs (n = 30), were collected. K. pneumoniae was isolated using lactose broth enrichment and MacConkey agar, with molecular confirmation via PCR targeting the gyrA and magA genes. PCR also identified ESBL genes (blaTEM, blaSHV, blaCTX-M, blaOXA-1) and biofilm genes (luxS, Uge, mrkD). Antimicrobial susceptibility was assessed, and the efficacy of five commercial disinfectants was evaluated by measuring inhibition zones. Klebsiella pneumoniae was isolated from poultry (13.3%), equine (8.3%), wild birds (15%), water (10%), feed (2%), and human hand swabs (6.6%). ESBL and biofilm genes were detected in the majority of the isolates, with significant phenotypic resistance to multiple antibiotics. The disinfectants containing peracetic acid and hydrogen peroxide were the most effective, producing the largest inhibition zones, while disinfectants based on sodium hypochlorite and isopropanol showed lower efficacy. Statistical analysis revealed significant differences in the effectiveness of disinfectants against K. pneumoniae isolates across various sample origins (P < 0.05). The presence of K. pneumoniae in animal and environmental sources, along with the high prevalence of ESBL-mediated resistance and biofilm-associated virulence genes, underscores the zoonotic potential of this pathogen. The study demonstrated that disinfectants containing peracetic acid and hydrogen peroxide are highly effective against ESBL-producing K. pneumoniae. Implementing appropriate biosecurity measures, including the use of effective disinfectants, is essential for controlling the spread of resistant pathogens in farm environments.

Bovine tracheal organoids for studying Mycoplasma bovis respiratory infections.

In vitro three-dimensional organoid models simulate key aspects of the structure and function of in vivo organs and have been used to study physiology, host-pathogen interactions, pathogenesis and pharmacodynamics. Although most organoid studies have been developed using human or mouse tissues, recent advancements have enabled the establishment of intestinal and respiratory tract organoids from domestic animal samples. Mycoplasma bovis causes chronic respiratory tract infections in cattle with significant health and economic consequences. The pathogenesis and virulence factors of M. bovis have been studied in several in vitro infection models, but the use of organoids has not been examined previously. In this study, we assessed the feasibility of using a matrix-embedded bovine tracheal organoid system to study respiratory infections with M. bovis. Bovine tracheal organoids were inoculated with M. bovis strain MbovMil and incubated for 72 hours to investigate the ability of M. bovis to proliferate, attach and invade the organoids. M. bovis was able to infect the organoids, resulting in a mean 260-fold increase in the titre of viable M. bovis by 72 hours post-inoculation. Examination of the infected organoids using transmission electron microscopy revealed the presence of mycoplasmas within the organoid cells and membrane bound clusters of M. bovis inside the intercellular junctions. Our findings indicate that bovine tracheal organoids can be used as a model system for studying respiratory tract infections caused by M. bovis.

Pengaruh Deterjen Cair terhadap Mortalitas dan Perilaku Ikan Nila (Oreochromis niloticus)

The disposal of detergent waste into water bodies leads to environmental problems. The use of detergents is increasing, and the variety of detergents available is also becoming more diverse. This study examines the mortality and behavior of Nile tilapia (Oreochromis niloticus) in response to liquid detergent contaminants. The research was conducted from March to April 2023 at the Fish Behaviour Laboratory, Faculty of Fisheries and Marine Sciences, IPB University. We used 154 individuals of Nile tilapia measuring 5-7 cm. A total of 90 individuals were used for the preliminary test and most of them died at the highest concentration of 100 ml/L within five minutes. Sixty individuals were used for determining LD50–96 and we found that the LD50–96 was 1.29 ml/L. The behavior of four individuals of tilapia was observed using focal animal sampling in four sublethal concentrations. The results show that the detergent contaminant inhibits oxygen diffusion and damages the physiological functions of the tilapia. The detergent concentration level also affects the operculum opening and closing behavior, movement, feeding response, and mortality.

Investigating the Welfare of Zoo-Housed Cryptoprocta ferox: Behavioral Observations and Hormonal Profiling.

The global decline of species necessitates intensive conservation efforts, including ex-situ breeding programs to safeguard endangered populations. However, managing welfare and reproduction in zoological gardens can present several challenges. This study aims to explore behaviors and endocrinological responses of two specimens of fossa (Cryptoprocta ferox) under human care. One hundred and twenty hours of observation through focal animal sampling continuous recording and 67 fecal samples were collected. From fecal samples, cortisol (FCM), progesterone (FPM), and testosterone (FTM) metabolites' concentrations were analyzed using Radio Immuno Assay. According to our results, higher FTM (p < 0.05) and FPM levels at the beginning of the study, showing a potential alignment with the breeding season. When the male entered the female outdoor area, he recorded longer durations of seeking and marking behaviors (p ≤ 0.01), along with lower FCM levels (p < 0.05), suggesting an enriching effect. In the same period, the female's FCM level was higher (p ≤ 0.01), indicating potential stress. When both animals were in their respective outdoor areas, the FCM level of the female remained significantly higher (p < 0.05), and the male showed a peak in pacing duration (p < 0.01), implying a potential stress or frustration status. In her indoor area, the female exhibited significantly shorter durations of locomotion, marking, and seeking (p < 0.01) and did not show any abnormal behavior, likely due to the reduced complexity of the environment. However, we cannot exclude that her complete visual and olfactory separation from the male and visitors may also have contributed to a decrease in her overall stress levels. The findings highlight the interplay between hormonal fluctuations and behavioral responses in the two zoo-housed fossa, providing essential insights for fostering their well-being and facilitating conservation efforts.

ИССЛЕДОВАНИЕ ВЛИЯНИЯ 20-ГИДРОКСИЭКДИЗОНА НА УГЛЕВОДНЫЙ ОБМЕН У ПОРОСЯТ

The effect of 20-hydroxyecdysone on carbohydrate metabolism indices was studied in crossbred piglets (♂ Danish Yorkshire ×♀ Danish Landrace) during the postnatal period of ontogenesis. At the age of 60 days, 2 groups of animals were formed: 1 (control, n=3) and 2 (experiment, n=4), which received compound feed con- taining (g/kg) crude protein 158.7; lysine 7.7; threonine 4.8; methionine 4.6 and metabolizable energy (MJ/kg) 12.7. 20-hydroxyecdysone was introduced into the diet of piglets in the experimental group at the rate of 30 mg/kg of feed. The parameters of carbohydrate metabolism (glucose, pyruvate, lactate concentrations, alpha-amylase and lactate dehydrogenase activity in the blood, glycogen content in the longissimus dorsi muscle and liver) were studied. Statistical processing of the obtained results was performed using the STATISTICA v.12.5 software package. The Mann-Whitney U-test was used to identify differences between animal samples. Differences be- tween groups were considered statistically significant at p≤0.05. The use of 20-hydroxyecdysone in animals dur- ing the postnatal period of ontogenesis for 2 months leads to a decrease in the lactate level in the blood serum (by 17.1%; p&lt;0.05). In animals of the experimental group, compared to the control, an increase in the pyruvate con- tent (by 15.8%; p&lt;0.05), the activity of lactate dehydrogenase and alpha-amylase enzymes in the blood serum was found against the background of a low glucose level in whole blood (by 22.8%; p&lt;0.05). At the same time, in an- imals receiving 20-hydroxyecdysone, compared to the control, an increase in the glycogen level in the liver and longissimus dorsi muscle was found (by 23.0% and 20.7%, respectively; p&lt;0.05). It was concluded that 20-hydroxyecdysone activates the intracellular signaling pathway of phosphatidylinositol-3-kinase and protein kinase B, which leads to an increase in the intensity and direction of carbohydrate metabolism, accompanied by complete oxidation of glucose to pyruvate with a large formation of adenosine triphosphoric acid (ATP) and ef- fective utilization of glucose for glycogen synthesis in the liver and longissimus dorsi muscle.

Ontogenetic change in body shape for white sharks, Carcharodon carcharias, in Australian waters.

The analysis of how biological shape changes across ontogeny can provide us with valuable information on how species adapt behaviorally, physiologically, and ecologically. The white shark Carcharodon carcharias is one of the largest and most widely distributed apex predators globally, yet an understanding of ontogenetic changes in body shape and relative scaling of length and weight measures is limited, especially in relation to foraging ecology. Through analysis of a suite of shape-related metrics, we identified ontogenetic patterns of scaling throughout development. Isometric growth was exhibited for most metrics, failing to show a significant deviation from an isometric slope of 1.0 for length-length relationships, and 3.0 for weight-length relationships. The most notable difference from this trend was the negative allometric growth observed for the upper caudal-fin lobe length, trunk length, and the mouth length. The surface area of the fins also presented a strong, positive relationship with precaudal length (PCL) and the girth at the pectoral fin. Negative allometric growth was exhibited for three of the fins (pectoral, upper caudal fin, and lower caudal fin) against PCL, exhibiting a significant deviation from the expected isometric growth of 2.0 for area-length relationships. There were no significant differences in morphometric relationships between geographic regions within Australia that samples were collected from. No differences between the sexes were identified; however, this may be an artifact of the lack of mature animal samples. Conversely, life stage was found to have a significant effect on the girth-length and weight-length relationships. The development of regression equations for morphometric measures allows the assessment of white shark body condition and may serve as an assessment tool to understand the potential impacts of human-induced environmental change on white sharks.

Comparison of the mid-infrared spectra and prediction equations developed from morning and evening milk samples from twice-a-day milked dairy cows.

Mid-infrared spectroscopy is a technology used globally for quantifying the concentration of fat, protein, lactose and other constituents in the milk samples of both individual animals and bulk tank milk. Differences in the milk components and yield of cows are known to exist between morning and evening milk; nonetheless, differences in the spectra originating from the same cow from morning and evening milkings have never been investigated. Data was obtained from 2,602 dairy cows from 7 research farms in Ireland. A total of 199,288 morning milk spectra with an associated evening milk spectra produced by the same cow within 24 h were available. Post editing, spectral data were available on the same 502 wavelengths in the mid-infrared region of the electromagnetic spectrum for all milk samples. Differences between morning and evening milk spectra produced by the same cow in a 24-h period were investigated using a) the mean and standard deviation of the difference between morning and evening spectra absorbance values, b) the correlation between the morning and the respective evening wavelength absorbance values, and c) the L2 distance all of which were quantified across 1) stages of lactation, 2) years, and 3) farms. The average (standard deviation of the) difference between the morning and the evening spectrum absorbance values produced by the same cow within 24 h was 0.00097 (0.008) and it was always larger than 0.055 for the wavelengths between 2,920 cm-1 and 2,947 cm-1. The correlation between morning wavelength absorbance values and the respective evening wavelength absorbance values were all strong (i.e., > 0.80) in the spectral region of 1,469 cm-1 to 1,473 cm-1; weak correlations of < 0.26 existed between morning and evening spectra wavelengths in the region of 1,593 cm-1 to 1,597 cm-1. These trends in correlations generally persisted within different stages of lactation, years, and farms. Results from the L2 distance indicated that early lactation morning and evening spectra were more different (P < 0.05) from each other than when compared in late lactation; no large differences in the L2 distance across different farms and years were evident. The impact of a prediction equation developed from morning spectral data but applied to evening milk spectral data, and vice versa, was investigated. Nitrogen use efficiency (NUE) was the trait explored which is a trait of the animal; the mean NUE (SD) in the validation data set was 22.16 (4.86). The root mean square error from predictions developed and validated on morning spectra samples was 3.49; this increased (P < 0.05) to 3.85 for the same validation in the morning spectra but when the prediction equation was developed using just evening spectra. Similarly, the root mean square error from predictions developed and validated on evening spectra samples was 3.46 which increased (P < 0.05) to 3.85 when the prediction equation was developed using just morning spectra. In conclusion, morning and evening milk spectra produced by the same cow within 24 h differ, particularly in some spectral regions; these differences impact the prediction performance of applied prediction equations.

Leptospira spp. is recognized by TLR2 and induces IFN-β and IFN-λ expression in smegma from naturally infected bulls.

Leptospirosis, a zoonosis with a wide worldwide distribution, causes significant reproductive losses in cattle. The genital presentation of the disease has been reported, and its understanding is crucial for the implementation of adequate sanitary measures. Although Leptospira spp. modulate innate immunity, the response in the genital mucosa of bulls is unknown. The objective of this work was to evaluate the expression of innate immune receptors and cytokines in smegma from bulls with genital infection by Leptospira spp. To do so, routine preputial scraping samples were selected from breeding bulls from four establishments in the Azul District, Buenos Aires, Argentina with no reproductive problems, which were previously evaluated for the presence of Leptospira spp. DNA by qPCR detection of the secY gene. The relative gene expression of TLR2, TLR4, IFN-β and IFN-λ in the smegma samples of animals infected with Leptospira spp. was determined by RT-qPCR, with subsequent analysis of the results using REST software. The expression of TLR2 was significantly increased in animals infected with Leptospira spp. compared to uninfected control animals (4-fold), while TLR4 did not show differences. Likewise, both IFN-β and IFN-λ were significantly increased in smegma of infected animals (3.5 and 3.1-fold, respectively). Therefore, this work shows that genital infection of Leptospira spp. in bulls generates and modulates a local innate immune response, with an association between the presence of Leptospira spp. DNA and the expression levels of TLR2, IFN-β and IFN-λ.

Ferroptosis is involved in the damage of ocular lens under long-term PM2.5 exposure in rat models and humans

Epidemiological studies show a positive association between air pollution and age-related cataracts, but the pathogenic mechanism remains unclear. This study first demonstrates that fine particulate matter (PM2.5) induces ferroptosis in the lens, leading to morphological and functional disorders, through human, animal, and cellular samples. In 3-week PM2.5-exposed rat models (10 µl 1 mg/ml PM2.5 suspension per eye, 4 times a day), we find that many vacuoles form in the lens equatorial region by analysis of haematoxylin and eosin staining after PM2.5 exposure. Using iron and glutathione (GSH) assay kits, we found increased Fe2+ contents and decreased GSH levels in PM2.5-exposed rats’ lenses. Additionally, the lipid peroxide 4-hydroxynonenal (4-HNE) was also found to be elevated with immunoblot, suggesting ferroptosis is involved. Ferroptosis was also observed in human lens epithelial cells treated with 25, 50, and 100 µg/ml PM2.5 suspension for 24 h, accompanied by decreased cell viability and migration. Furthermore, we collect about 60 human lens anterior capsule (HLAC) samples for RNA-seq. The results show that compared to HLACs from areas with PM2.5 concentration ≤30 μg/m³, ferroptosis-related genes expression of those from areas with PM2.5 concentration ≥35 μg/m³ are significantly altered, such as glutathione peroxidase 4 and STEAP family member 3. Also, human lens in areas with high PM2.5 concentrations showed elevated levels of transferrin receptor and 4-HNE with immunoblot, and down-regulated expression of connexin 43 (Cx43) through immunofluorescent. These results demonstrate that ferroptosis plays a key role in PM2.5-induced cataractogenesis.

Cryptosporidium Species Infections Detected from Fecal Samples of Animal and Human Hosts in South Africa: Systematic Review and Meta-Analysis.

This study presents a systematic review and meta-analysis approach of Cryptosporidium species prevalence studies in animal and human hosts published between 1980 and 2020 in South Africa. Extensive searches were conducted on three electronic databases including PubMed, ScienceDirect and Google Scholar. The findings indicated an overall pooled prevalence estimate (PPE) of Cryptosporidium spp. infections in animals and humans at 21.5% and 18.1%, respectively. The PCR-RFLP appeared to be the most sensitive diagnostic method with a PPE of 77.8% for the detection of Cryptosporidium spp. infections followed by ELISA (66.7%); LAMP (45.4%); PCR (25.3%); qPCR (20.7%); microscopy (10.1%); IFAT (8.4%); and RDT (7.9%). In animal hosts, C. parvum had the highest PPE of 3.7%, followed by C. andersoni (1.5%), C. ubiquitum (1.4%) and C. bovis (1.0%), while in humans, C. parvum also had the highest PPE of 18.3% followed by C. meleagridis at 0.4%. The data generated in this study indicated that Cryptosporidium spp. infections were highly prevalent in both animals and humans in South Africa, especially in the KwaZulu-Natal and North West provinces. However, we further observed that there was a lack of prevalence studies for both animals and humans in some of the provinces. This study highlights the necessity for a "One Health" strategic approach promoting public hygiene, animal husbandry and regular screening for Cryptosporidium spp. infections in both animals and humans.

The Development of a High-Throughput Homonuclear Decoupling HSQC NMR Platform for the Determination of 10 Sex Hormones in Animal-Source Food and Medicines

Owing to their endocrine disruption effect, the hormone levels in animal-source food and medicines need to be efficiently and accurately quantified by a reliable analytical method. In the current study, by using a homonuclear decoupling and heteronuclear single quantum correlation (HSQC) experiment, coupled with non-uniform sampling (NUS) that was used to shorten the experimental time, we developed a method to quantify 10 hormone residues in animal-source products. This method was validated following the guidelines of USP–NF 2022. The application of the homonuclear decoupling (HD) technique to conventional HSQC yielded 2D spectra that exhibited excellent signal separation and specificity. For all the tested hormones, good linearity with correlation coefficients of more than 0.99 was observed in the linear range of 0.2–6 mg/0.6 mL. Satisfactory precision and recoveries of spiked animal samples were also obtained. Finally, the method was applied in residue determination of 10 hormones in real animal-source samples at the ug/g level.

Dehydroascorbic acid quantification in human plasma: Simultaneous direct measurement of the ascorbic acid/dehydroascorbic acid couple by UPLC/MS-MS

Ascorbic acid (AA, vitamin C) and dehydroascorbic acid (DHA) constitute a biological couple. No technique can accurately, independently, and simultaneously quantify both members of the couple in animal and human samples, thereby constraining advances in physiology and pathophysiology. Here we describe a new UPLC/MS/MS method to measure both compounds directly and independently in human plasma. Lower limits of quantification were 16 nM, with linear coefficients >0.99 over a 100-fold concentration range. The method was stable and reproducible with <10 % injection-to-injection variation. Use of isotopic labeled internal standards for both compounds ensured precision and accuracy. Plasma preparation required only 2 steps. In plasma samples from 14 anonymized subjects who met criteria for blood donation, mean concentrations were 6±2 μmol/L (mean ± SD) and 56 ± 14 μmol/L for DHA and AA respectively, with (DHA)/(AA + DHA) ratio of 9.8 %. This method represents a pioneering approach to measuring the AA/DHA couple in human plasma.

Occurrence and assemblage distribution of Giardia Duodenalis in symptomatic and asymptomatic patients in southeastern Iran (2019–2022)

BackgroundThe ubiquitous protozoan parasite Giardia duodenalis is a major contributor to the global burden of diarrhoea, particularly in young children living in poor-resource regions. Although rarely mortal, giardiasis is associated with growth retardation and cognitive impairment in early childhood. Here we investigate the epidemiology of human giardiasis in Iranshahr (south-eastern Iran), a region where this information was previously lacking.MethodsStool samples were collected from 17,455 outpatients and inpatients attended at three major hospital settings during April 2020 and March 2022. Microscopy was used as a screening method for the presence of Giardia cysts, and the identification of G. duodenalis assemblages was carried out using PCR and Sanger sequencing.ResultsThe overall prevalence of giardiasis was 1.87 (326/17,455; 95% CI: 1.7–2.1). Being female was positively associated with higher odds of giardiasis (p = 0.014). Individuals without diarrhoea were less likely to have giardiasis (p = 0.022). Individuals attending the Iran Hospital were more likely to harbour G. duodenalis infections compared to those attending at the Khatam Hospital and the Clinical Reference Laboratory (p = 0.001). Our sequence analyses revealed the presence of assemblages A (56.5%, 13/23), B (39.1%, 9/23), and A + B (4.4%, 1/23). No association was observed between the occurrence of a given assemblage and the occurrence of diarhroea.ConclusionsGiardia infections were found at relatively low prevalence rates in both symptomatic and asymptomatic individuals seeking medical attention. Being female, having diarrhoea, and being sampled during 2020–21 were predictors of giardiasis. Although limited, our molecular data indicate that some Giardia infections may be zoonotic in nature. These data should be corroborated and expanded in future epidemiological studies targeting simultaneously human, animal, and environmental (water) samples to improve our understanding of the epidemiology of giardiasis in Iran.

Natural Feed Options for Sumatran Elephants at the Seblat Elephant Training Center North Bengkulu Regency, Indonesia

The health and welfare of Sumatran elephants at the Seblat Elephant Training Center (SETC), North Bengkulu Regency, depend on many environmental factors in their habitat, one of which is the availability of a variety of plant species (diet) in the Seblat Nature Tourism Park (SNTP) forest area. Research on this matter was conducted in May-June 2022 within the SNTP forest area, where the Seblat SETC facility is located. This study aimed to evaluate the dietary preferences of Sumatran elephants at SETC towards natural plant species in the SNTP forest. The "Focal Animal Sampling" method was used to measure the duration of feeding activity of each elephant on specific plant species while they were foraging in the forest. The elephants' preferences for plant species were analyzed quantitatively, while the types of plants consumed by the elephants were described qualitatively. The research found that 15 families with 33 species were identified as the natural food plants for the SETC elephants in Bengkulu. The Poaceae family was most preferred by the elephants with the highest preference value (35.07%), followed by Fabaceae (31.73%), Malvaceae (9.87%), and Cyperaceae (7.83%), while the least preferred were Costaceae, and Myrtaceae (0.33% respectively), Convolvulaceae (0.41%), and Selaginellaceae (0.73%). Based on sex, adult male elephants preferred Fabaceae plants such as Mimosa pudica (35.99%), Desmodium triflorum (11.66%), Mimosa invisa (7.36%), Cynodon dactylon (6.54%) over other families. Conversely, adult female elephants preferred Poaceae such as Leersia virginica Willd., Cyrtococcum patens (L.) A., Centhotheca lappacea Desvaux., Digitaria sanguinalis (L.) Scop., and Imperata cylindrica L. patients. These predictors, however, need further work to validate reliability.

The Effect of Giving Soursop Fruit Juice (Annona muricata, L.) on Hemoglobin Levels in Wistar Bunting Strain Rats Exposed to Cigarette Smoke

Background: Anemia in pregnancy can be caused by exposure to cigarette. A large number of free radicals can cause the number of erythrocytes to decrease. It takes a component to neutralize free radicals, namely antioxidants obtained from various fruits. Soursop contains sources of antioxidants such as flavonoid compounds, polyphenols, and vitamin C. Researchers want to know whether soursop juice can be used as an alternative antioxidant to prevent the decrease in hemoglobin levels of pregnant women. Objective: To determine the effect of giving soursop juice on hemoglobin levels of pregnant white rats with exposure to cigarette smoke. Methods: The experimental laboratory research was designed using the Randomized Post Test Only Control Group Design method. The experimental animal samples were 20 pregnant female Wistar (Rattus norvegicus) rats composed of two control and three treatment groups. Rats were given soursop fruit juice from the first day of rearing until the 18th day of gestation and exposed to cigarette smoke from the 6th day until the 18th. Then blood samples were taken from the mother's heart to measure the hemoglobin level. Results: The results of the hemoglobin level test showed that there was difference between the treatment group and control group. There is an increasing trend from the postitive control group to treatment group. Conclusion: Giving soursop fruit juice can prevent the decrease in hemoglobin levels of pregnant rats exposed to cigarette smoke. Keywords: anemia, free radicals, antioxidants, vitamin C.

Does Every Strain of Pseudomonas aeruginosa Attack the Same? Results of a Study of the Prevalence of Virulence Factors of Strains Obtained from Different Animal Species in Northeastern Poland.

Pseudomonas aeruginosa is a pathogen that causes infections in animals and humans, with veterinary implications including ear infections in dogs, respiratory diseases in cats, and mastitis in ruminants. In humans, it causes severe hospital-acquired infections, particularly in immunosuppressed patients. This study aimed to identify and assess the prevalence of specific virulence factors in Pseudomonas aeruginosa isolates. We analyzed 98 Pseudomonas aeruginosa isolates from various animal samples (dogs, cats, ruminants, fowl) from northeastern Poland in 2019-2022 for virulence-related genes (toxA, exoU, exoT, exoS, lasB, plcN, plcH, pldA, aprA, gacA, algD, pelA, endA, and oprF) by PCR and assessed biofilm formation at 48 and 72 h. Genomic diversity was assessed by ERIC-PCR. The obtained results showed that all strains harbored the pelA gene (100%), while the lowest prevalence was found for pldA (24%) and exoU (36%). Regardless of the animal species, strong biofilm forming ability was prevalent among the strains after both 48 h (75%) and 72 h (74%). We obtained as many as 87 different genotyping profiles, where the dominant one was profile ERIC-48, observed in four strains. No correlation was found between presence or absence of determined genes and the nature of infection. Similarly, no correlation was found between biofilm-forming genes and biofilm strength. The high genetic diversity indicates challenges for effective prevention, emphasizing the need for ongoing monitoring and research.

Design characteristics of integrated surveillance of antimicrobial resistance at the human, animal, and environment interfaces

Abstract Efforts have been made to strengthen national and global antimicrobial resistance (AMR) surveillance programs by integrating data collection across the human, animal, and environmental sectors. We searched the literature to review published studies reporting the implementation of integrated AMR surveillance approaches, and we identified a total of 96 articles from 36 countries published from 2000 to 2022, which met our inclusion criteria. Standard review protocols were applied in our study. Out of 96 studies, 47 (49%) articles integrated AMR analysis from human and animal (HA) populations, 24 (25%) considered human, animal, and environmental (HAE) samples together, 11 (12%) studies included animal and environment (AE) samples, and 12 (13%) integrated human and environmental (HE) surveillance. Human isolate-based and animal isolate-based surveillance were the most common study designs (38, 52.8%), with the remainder applying human case-based and isolate-based for animal populations (32, 44.4%). Finally, our results demonstrate that AMR studies on E. coli revealed a lower correlation between human and animal AMR prevalence compared to Salmonella spp. and Campylobacter spp. studies. Different aspects of surveillance design were associated with the level of correlation of AMR prevalence between sectors. Our study found that while global efforts for integrated AMR surveillance have increased in the past 10 years, significant variation exists between studies with regard to the epidemiological and laboratory aspects of their surveillance designs. Our findings indicate that to enable the generation of comparable epidemiological data across countries and sectors, there is a need for the development of a global protocol to support the design of surveillance programs that aim to conduct integrated surveillance of AMR.