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Pen-based oral fluids are used extensively for surveillance and disease detection in swine, but there is sparse information on the sampling process itself. To address this shortcoming, we documented the pen-based oral fluid sampling process with the aim of optimizing the number of pigs in a pen that contributed to the sample. We quantified the effects of (1) previous experience with rope sampling (training), (2) the number of ropes suspended in the pen, and (3) sampling time on pig participation and pig-rope contact. A subset of pigs was clearly marked for individual identification and their interactions with ropes video recorded. Thereafter, pig-rope contacts were counted from the recordings, with "contact" defined as an individually identified pig clearly taking the rope into its mouth. Data were analyzed using appropriate models (R version 4.4.1 R core team 2024). Training, provision of additional ropes, and extended sampling time all increased pig participation across pen sizes. However, for routine oral fluid collection in the field, we recommend training pigs prior to hanging ropes for sample collection and increasing sampling time to maximize the pigs' contribution to the oral fluid sample. Importantly, these studies focused on pig behavior and not detection; thus, future studies should evaluate the impact of these same factors on the probability of detection. [ABSTRACT FROM AUTHOR]

Objectives: This study investigated the dynamics of porcine circovirus type 2 (PCV2) and PCV3 on a commercial farm following PCV2 vaccination. Methods: Serum samples from 35 pigs, starting at 3 weeks of age, were collected weekly until 21 weeks of age. Oral fluids from six pens of pigs of the same age were also analyzed. Viral DNA was assessed in pooled sera and individual oral fluid samples, while antibodies (IgG and IgA) were measured in the serum and oral fluids. Productive parameters, including weekly mortality and cumulative mortality, were evaluated. Results: The results revealed that PCV2 and PCV3 co-infection was detected in pigs at 8 weeks of age, with PCV3 being detected in oral fluids two weeks earlier. PCV3 DNA was detected in oral fluids at 4 weeks of age. PCV2 IgG antibodies in the serum increased gradually after vaccination, peaking at 7 weeks of age, then declined and stabilized until 21 weeks of age. PCV3 IgG antibodies fluctuated early but were uniformly positive after 13 weeks of age. In oral fluids, PCV2 IgG and IgA antibodies showed a strong response only at 3 and 23 weeks of age. In contrast, a strong and consistent IgG response was observed in oral fluids in the absence of PCV2 and PCV3 co-infection of pigs at 3 to 11 weeks of age. The farm's productive parameters remained stable throughout the study. Conclusions: These findings suggest that PCV2 and PCV3 co-infection, along with high PCV3 detection levels in serum and oral fluids, may have an impact on the efficacy of PCV2 vaccination. [ABSTRACT FROM AUTHOR]

In recent years, synthetic cannabinoids (SCs) have become a major public health issue. For this reason, there is a need for innovative analytical methods that allow its monitoring in biological matrices. In this work, we propose a novel methodology to screen eight SCs (AM-694, cumyl-5F-PINACA, MAM-2201, 5F-UR-144, JWH-018, JWH-122, UR-144 and APINACA) in oral fluids. A bar adsorptive microextraction method followed by microliquid desorption combined with high-performance liquid chromatography with diode array detection (BAµE-µLD/HPLC-DAD) was developed to monitor the target SCs. The main factors affecting the BAµE technology were fully optimized for oral fluid analysis. Under optimized experimental conditions, the proposed methodology showed good linear dynamic ranges from 20.0 to 2000.0 µg L−1 (r2 > 0.99, relative residuals < 15%), limits of detection between 2.0 and 5.0 µg L−1 and suitable average recovery yields (87.9–100.5%) for the eight studied SCs. The intra- and interday accuracies (bias ≤ ± 14.7%) and precisions (RSD ≤ 14.9%) were also evaluated at three spiking levels. The validated methodology was then assayed to oral fluid samples collected from several volunteers. The proposed analytical approach showed remarkable performance and could be an effective alternative for routine monitoring of the target compounds in oral fluid. [ABSTRACT FROM AUTHOR]

The objective of this study is to analyze the miRNA expression of oral fluids such as gingival crevicular fluid (GCF) in patients with periodontitis and Type 2 diabetes mellitus, and how these epigenetic biomarkers can influence the bidirectional relationship of these two inflammatory diseases. This review was conducted following the PRISMA criteria. PubMed, Scopus, Cochrane Library, Embase, and Web of Science databases were searched for clinical studies conducted on humans investigating, through GCF miRNA expression, the relationship between periodontal diseases and type 2 diabetes mellitus. In addition, the etiopathogenic pathways of the studied miRNAs were analyzed using the DIANA MIR path tool. A total of 1436 references were identified in the initial literature search, and seven articles were finally included in this review. Most of the articles included in this review were case–control studies and examined the expression of miRNAs in patients with periodontitis with or without diabetes. Due to their characteristics, miRNAs appear to be the ideal biomarkers for improving the understanding and knowledge of the etiopathogenic pathways that link both diseases. Among all the studied miRNAs, miR-146a, miR-155, miR-200b, miR-223, and miR-203 showed strong involvement in inflammatory and metabolic pathways, making them potential good diagnostic and prognostic biomarkers. [ABSTRACT FROM AUTHOR]

A 24/7 AI sound-based coughing monitoring system was applied in combination with oral fluids (OFs) and bioaerosol (AS)-based screening for respiratory pathogens in a conventional pig nursery. The objective was to assess the additional value of the AI to identify disease patterns in association with molecular diagnostics to gain information on the etiology of respiratory distress in a multimicrobially infected pig population. Respiratory distress was measured 24/7 by the AI and compared to human observations. Screening for swine influenza A virus (swIAV), porcine reproductive and respiratory disease virus (PRRSV), Mycoplasma (M.) hyopneumoniae, Actinobacillus (A.) pleuropneumoniae, and porcine circovirus 2 (PCV2) was conducted using qPCR. Except for M. hyopneumoniae, all of the investigated pathogens were detected within the study period. High swIAV-RNA loads in OFs and AS were significantly associated with a decrease in respiratory health, expressed by a respiratory health score calculated by the AI The odds of detecting PRRSV or A. pleuropneumoniae were significantly higher for OFs compared to AS. qPCR examinations of OFs revealed significantly lower Ct-values for swIAV and A. pleuropneumoniae compared to AS. In addition to acting as an early warning system, AI gained respiratory health data combined with laboratory diagnostics, can indicate the etiology of respiratory distress. [ABSTRACT FROM AUTHOR]

Swine viral diseases have the capacity to cause significant losses and affect the sector's sustainability, a situation further exacerbated by the lack of antiviral drugs and the limited availability of effective vaccines. In this context, a novel point-of-care (POC) diagnostic device incorporating photonic integrated circuits (PICs), microfluidics and information, and communication technology into a single platform was developed for the field diagnosis of African swine fever (ASF) and classical swine fever (CSF). The device targets viral particles and has been validated using oral fluid and serum samples. Sensitivity, specificity, accuracy, precision, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR) were calculated to assess the performance of the device, and PCR was the reference method employed. Its sensitivities were 80.97% and 79%, specificities were 88.46% and 79.07%, and DOR values were 32.25 and 14.21 for ASF and CSF, respectively. The proposed POC device and PIC sensors can be employed for the pen-side detection of ASF and CSF, thus introducing novel technological advancements in the field of animal diagnostics. The need for proper validation studies of POC devices is highlighted to optimize animal biosecurity. [ABSTRACT FROM AUTHOR]

Pen-based oral fluids are used extensively for surveillance and disease detection in swine, but there is sparse information on the sampling process itself. To address this shortcoming, we documented the pen-based oral fluid sampling process with the aim of optimizing the number of pigs in a pen that contributed to the sample. We quantified the effects of (1) previous experience with rope sampling (training), (2) the number of ropes suspended in the pen, and (3) sampling time on pig participation and pig-rope contact. A subset of pigs was clearly marked for individual identification and their interactions with ropes video recorded. Thereafter, pig-rope contacts were counted from the recordings, with “contact” defined as an individually identified pig clearly taking the rope into its mouth. Data were analyzed using appropriate models (R version 4.4.1 R core team 2024). Training, provision of additional ropes, and extended sampling time all increased pig participation across pen sizes. However, for routine oral fluid collection in the field, we recommend training pigs prior to hanging ropes for sample collection and increasing sampling time to maximize the pigs’ contribution to the oral fluid sample. Importantly, these studies focused on pig behavior and not detection; thus, future studies should evaluate the impact of these same factors on the probability of detection.

Objectives: This study investigated the dynamics of porcine circovirus type 2 (PCV2) and PCV3 on a commercial farm following PCV2 vaccination. Methods: Serum samples from 35 pigs, starting at 3 weeks of age, were collected weekly until 21 weeks of age. Oral fluids from six pens of pigs of the same age were also analyzed. Viral DNA was assessed in pooled sera and individual oral fluid samples, while antibodies (IgG and IgA) were measured in the serum and oral fluids. Productive parameters, including weekly mortality and cumulative mortality, were evaluated. Results: The results revealed that PCV2 and PCV3 co-infection was detected in pigs at 8 weeks of age, with PCV3 being detected in oral fluids two weeks earlier. PCV3 DNA was detected in oral fluids at 4 weeks of age. PCV2 IgG antibodies in the serum increased gradually after vaccination, peaking at 7 weeks of age, then declined and stabilized until 21 weeks of age. PCV3 IgG antibodies fluctuated early but were uniformly positive after 13 weeks of age. In oral fluids, PCV2 IgG and IgA antibodies showed a strong response only at 3 and 23 weeks of age. In contrast, a strong and consistent IgG response was observed in oral fluids in the absence of PCV2 and PCV3 co-infection of pigs at 3 to 11 weeks of age. The farm’s productive parameters remained stable throughout the study. Conclusions: These findings suggest that PCV2 and PCV3 co-infection, along with high PCV3 detection levels in serum and oral fluids, may have an impact on the efficacy of PCV2 vaccination.

Simple Summary: The transfer of a target directly from pigs or indirectly from the environment into pen-based oral fluid samples was documented using a fluorescent tracer (red food coloring). Pens of ~30, ~60, and ~125 14-week-old pigs (32 pens/size) on commercial swine farms received one of two treatments: (1) pig exposure, i.e., ~3.5 mL of tracer solution sprayed into the mouth of 10% of the pigs in the pen; (2) environmental exposure, i.e., 20 mL of tracer solution was poured on the floor in the center of the pen. Oral fluids collected one day prior to treatment (baseline fluorescence control) and immediately after treatment were tested for fluorescence. Data were evaluated by receiver operating characteristic (ROC) analysis, with Youden's J statistic used to set a threshold. Pretreatment oral fluid samples with fluorescence responses above the ROC threshold were removed from further analysis (7 of 96 samples). Analysis of post-treatment samples showed detectable levels of red food coloring in oral fluid samples from 78 of 89 pens (87.6%), including 43 of 47 (91.5%) pens in which pigs were directly exposed and 35 of 42 (83.3%) pens in which the tracer was placed in the environment. Thus, oral fluid samples contain both pig-derived and environmental targets. Laboratory methods for detecting specific pathogens in oral fluids are widely reported, but there is little research on the oral fluid sampling process itself. In this study, a fluorescent tracer (diluted red food coloring) was used to test the transfer of a target directly from pigs or indirectly from the environment to pen-based oral fluid samples. Pens of ~30, ~60, and ~125 14-week-old pigs (32 pens/size) on commercial swine farms received one of two treatments: (1) pig exposure, i.e., ~3.5 mL of tracer solution sprayed into the mouth of 10% of the pigs in the pen; (2) environmental exposure, i.e., 20 mL of tracer solution was poured on the floor in the center of the pen. Oral fluids collected one day prior to treatment (baseline fluorescence control) and immediately after treatment were tested for fluorescence. Data were evaluated by receiver operating characteristic (ROC) analysis, with Youden's J statistic used to set a threshold. Pretreatment oral fluid samples with fluorescence responses above the ROC threshold were removed from further analysis (7 of 96 samples). Based on the ROC analyses, oral fluid samples from 78 of 89 pens (87.6%), contained red food coloring, including 43 of 47 (91.5%) pens receiving pig exposure and 35 of 42 (83.3%) pens receiving environmental exposure. Thus, oral fluid samples contain both pig-derived and environmental targets. This methodology provides a safe and quantifiable method to evaluate oral fluid sampling vis-à-vis pen behavior, pen size, sampling protocol, and target distribution in the pen. [ABSTRACT FROM AUTHOR]

The early detection of classical swine fever (CSF) remains a key challenge, especially when outbreaks are caused by moderate and low-virulent CSF virus (CSFV) strains. Oral fluid is a reliable and cost-effective sample type that is regularly surveilled for endemic diseases in commercial pig herds in North America. Here, we explored the possibility of utilizing oral fluids for the early detection of CSFV incursions in commercial-size pig pens using two independent experiments. In the first experiment, a seeder pig infected with the moderately-virulent CSFV Pinillos strain was used, and in the second experiment, a seeder pig infected with the highly-virulent CSFV Koslov strain was used. Pen-based oral fluid samples were collected daily and individual samples (whole blood, swabs) every other day. All samples were tested by a CSFV-specific real-time RT-PCR assay. CSFV genomic material was detected in oral fluids on the seventh and fourth day post-introduction of the seeder pig into the pen, in the first and second experiments, respectively. In both experiments, oral fluids tested positive before the contact pigs developed viremia, and with no apparent sick pigs in the pen. These results indicate that pen-based oral fluids are a reliable and convenient sample type for the early detection of CSF, and therefore, can be used to supplement the ongoing CSF surveillance activities in North America. [ABSTRACT FROM AUTHOR]

Aim: To determine the accuracy of biomarker combinations in gingival crevicular fluid (GCF) and saliva through meta‐analysis to diagnose periodontitis in systemically healthy subjects. Methods: Studies on combining two or more biomarkers providing a binary classification table, sensitivity/specificity values or group sizes in subjects diagnosed with periodontitis were included. The search was performed in August 2022 through PUBMED, EMBASE, Cochrane, LILACS, SCOPUS and Web of Science. The methodological quality of the articles selected was evaluated using the QUADAS‐2 checklist. Hierarchical summary receiver operating characteristic modelling was employed to perform the meta‐analyses (CRD42020175021). Results: Twenty‐one combinations in GCF and 47 in saliva were evaluated. Meta‐analyses were possible for six salivary combinations (median sensitivity/specificity values): IL‐6 with MMP‐8 (86.2%/80.5%); IL‐1β with IL‐6 (83.0%/83.7%); IL‐1β with MMP‐8 (82.7%/80.8%); MIP‐1α with MMP‐8 (71.0%/75.6%); IL‐1β, IL‐6 and MMP‐8 (81.8%/84.3%); and IL‐1β, IL‐6, MIP‐1α and MMP‐8 (76.6%/79.7%). Conclusions: Two‐biomarker combinations in oral fluids show high diagnostic accuracy for periodontitis, which is not substantially improved by incorporating more biomarkers. In saliva, the dual combinations of IL‐1β, IL‐6 and MMP‐8 have an excellent ability to detect periodontitis and a good capacity to detect non‐periodontitis. Because of the limited number of biomarker combinations evaluated, further research is required to corroborate these observations. [ABSTRACT FROM AUTHOR]

Simple Summary: Multiple viruses and bacteria can cause respiratory disease in pigs. We aimed to report how frequently certain viruses and bacteria were detected in samples from pigs with respiratory disease in the course of routine diagnostic procedures at the University of Veterinary Medicine in Vienna between 2016 and 2021. While Mycoplasma (M.) hyorhinis (55.1%) had the highest detection rate, influenza A virus had the lowest detection rate (6.1%) in the investigated samples. Lung samples tested positive for PRRSV RNA were also more likely to be positive for M. hyopneumoniae and Pasteurella (P.) multocida. Samples tested positive for M. hyopneumoniae were more likely to be positive for P. multocida and Streptococcus suis, but less likely to be positive for M. hyorhinis. In conclusion, lung samples that were positive for a primary pathogenic agent were more likely to be positive for a secondary pathogenic agent. The diagnostic workup of respiratory disease in pigs is complex due to coinfections and non-infectious causes. The detection of pathogens associated with respiratory disease is a pivotal part of the diagnostic workup for respiratory disease. We aimed to report how frequently certain viruses and bacteria were detected in samples from pigs with respiratory symptoms in the course of routine diagnostic procedures. Altogether, 1975 routine diagnostic samples from pigs in Austrian swine stocks between 2016 and 2021 were analysed. PCR was performed to detect various pathogens, including porcine reproductive and respiratory syndrome virus (PRRSV) (n = 921), influenza A virus (n = 479), porcine circovirus type 2 (PCV2) (n = 518), Mycoplasma (M.) hyopneumoniae (n = 713), Actinobacillus pleuropneumoniae (n = 198), Glaesserella (G.) parasuis (n = 165) and M. hyorhinis (n = 180). M. hyorhinis (55.1%) had the highest detection rate, followed by PCV2 (38.0%) and Streptococcus (S.) suis (30.6%). PRRSV was detected most frequently in a pool of lung, tonsil and tracheobronchial lymph node (36.2%). G. parasuis was isolated more frequently from samples taken after euthanasia compared to field samples. PRRSV-positive samples were more likely to be positive for PCV2 (p = 0.001), M. hyopneumoniae (p = 0.032) and Pasteurella multocida (p < 0.001). M. hyopneumoniae-positive samples were more likely to be positive for P. multocida (p < 0.001) and S. suis (p = 0.046), but less likely for M. hyorhinis (p = 0.004). In conclusion, our data provide evidence that lung samples that were positive for a primary pathogenic agent were more likely to be positive for a secondary pathogenic agent. [ABSTRACT FROM AUTHOR]

A fast and simple procedure based on microextraction by packed sorbent (MEPS) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been developed for the simultaneous quantification of 28 synthetic hallucinogens in oral fluids, including lysergic acid diethylamide and substances from NBOMe, NBOH, NBF, 2C, and substituted amphetamine categories. Extraction conditions such as type of sorbent, sample pH, number of charge/discharge cycles, and elution volume were studied. Hallucinogenic compounds were extracted from oral fluid samples using C18 MEPS, loading with 100 μL sample (adjusted to pH 7) in 3 cycles, washing with 100 μL deionized water, and eluting with 50 μL methanol in 1 cycle, giving quantitative recoveries and no significant matrix effects. Limits of detection from 0.09 to 1.22 μg L−1; recoveries from 80 to 129% performed in spiked oral fluid samples at 20, 50, and 100 μg L−1; and high precision with relative standard deviations lower than 9% were obtained. The proposed methodology was demonstrated to be appropriate for the simple and sensitive determination of NBOMe derivates and other synthetic hallucinogenic substances in oral fluid samples. [ABSTRACT FROM AUTHOR]

Aim: The intraoral scanners are digital devices used to digitise the oral tissues. The accuracy of the intraoral scanners has been studied under different environmental conditions, but there might be differences that occur in the actual oral environment, which is still in question. The aim of the study was to evaluate the accuracy and efficiency of Parallel Confocal Microscopy and 3D in motion video with triangulation technologybased intraoral scanners under the influence of moisture and mouth opening. Settings and Design: This was an Cross over clinical controlled study. Materials and Methods: The controlled in vivo study included healthy subjects who were in need of CBCT for the purpose of locating the position of unerupted third molars before going abroad for a job. The subjects were exposed to scans in the upper and lower jaws with two intraoral scanners based on 3D motion video technology with triangulation (Medit) and parallel confocal microscopy (Trios) under the influence of two oral conditions, which were moisture (presence and absence of moisture) and mouth opening (30 mm and 50 mm, respectively). A total of 96 scans were obtained and superimposed individually over the reference CBCT scans to find the deviations in the Geomagic Rapidform (version 2020, USA) software. The efficiency of the scanners was calculated by recording the time taken and the number of images obtained after each scan. Statistical Analysis Used: The significance was calculated by using the independent and paired sample t test in SPSS software (IBM, version 23). Results: Based on the surface analysis, the trueness of the intra-oral scanners had statistically significant differences when compared between 3D in motion video technology with Triangulation and Parallel Confocal Microscopy (P < 0.05) whereas no statistical significance was observed in precision. There was a significant difference observed in the efficiency of the intra-oral scanners (P < 0.05). Conclusion: There is a significant difference in the accuracy and efficiency of the intraoral scanners under the influence of oral conditions, such as different moisture levels and mouth opening conditions. 3D in motion video technology with Triangulation showed better results with the least deviation than Parallel Confocal Microscopy. [ABSTRACT FROM AUTHOR]

Abstract Background According to the World Health Organization, road traffic injuries lead to 1.3 million deaths each year and represent the leading cause of death for young adults under 30 years old. The use of psychoactive substances, including alcohol, drugs and pharmaceuticals, is a well-known risk factor for road traffic injuries. Our study aims to assess the prevalence of substances consumed by drivers in western Switzerland. Such studies are pivotal to improving prevention and developing public awareness campaigns. Methods To assess the prevalence of psychoactive substances among drivers, roadside controls were performed in collaboration with local police, using their classical sampling procedures to detect drivers under the influence of drugs or alcohol over two time periods (P1: 2006-2008, P2: 2017-2020). When impaired driving was not suspected by the police, minimally invasive sampling strategies (i.e., oral fluids during P1 and dried blood spots during P2) were performed on volunteer drivers after a road safety survey. A posteriori analyses and statistical interpretation were then performed. Results Among the 1605 drivers included in the study, 1048 volunteers provided an oral fluid sample, while 299 provided a dried blood spot sample. The percentage of drivers testing positive for at least one substance that can impact driving abilities was stable over time, with a rate of 10.5% positivity measured over both periods. Considering the different categories of substances, a slight variation was observed between both periods, with 7.6 and 6.3% of pharmaceuticals and 3.6 and 4.9% of illicit drugs for P1 and P2, respectively. Regarding the consumption of illicit drugs, the highest percentage of positivity was measured in biological fluids of drivers under the age of 35, during nights and week-ends, periods which are considered particularly prone to fatal accidents for this age group. Disturbingly, the road safety survey highlighted that drivers’ perception of the risk of getting positively controlled while driving after drug consumption is low (3.3 on a 1-to-10 scale, N = 299). Conclusion The number of positive cases measured in voluntary drivers who passed the preliminary police check demonstrates the importance of systematic biofluid sampling strategies regarding driving under the influence of psychoactive substances. Although the number of fatal road accidents globally has decreased over time, the results of this study reveal the need for both better prevention and deterrent processes that could potentially reduce the risk of fatal road accidents associated with drug consumption.

In this manuscript, a paper-based device modified with metal-organic frameworks was applied for the extraction of synthetic cannabinoid receptor agonists (SCRAs) in oral fluids, taking profit of the several interaction mechanisms between MOF-analytes, followed by their quantification by LC-fluorescence detection. The resulting composite, named as NH2-UiO-66@paper, was characterized by different techniques such as infrared, scanning electron microscopy, among others. The extraction procedure was optimized by using a full factorial design after an appropriate selection of the significant variables. Once the method was optimized, the analytical parameters were studied including liner range (LOQ-400 µg L−1), limit of detection (0.7 to 2.8 µg L−1) and intra- and inter-batch precision (

Background: Influenza A virus (IAV) surveillance in swine is critical not only due to the direct impact of the disease in the pork industry but also because IAV are prone to interspecies transmission (from human to pigs and vice versa); therefore, its monitoring is fundamental from a public and animal health perspective. Several diagnostic techniques have been used to detect IAV infection from nasal samples in swine, while samples of oral fluids (OF) are in use as novel alternatives for pathogen detection. The OF allow for efficient and feasible low-cost disease detection at the herd level, with low risk of stress for the animals. Objective: To describe a surveillance strategy of IAV at the herd level during respiratory disease outbreaks in swine farms at tropical settings using porcine oral fluids. Methods: An active surveillance strategy was conducted in several farms with past records of respiratory disease. The IAV detection was conducted in five purposively selected swine farms from years 2014 to 2017. We investigated a total of 18 respiratory outbreaks of the disease. Swine OF were collected for IAV testing. An OF sample is described as a pen-based specimen collected from a group of >20 pigs per pen and/or per barn (stall-housed individually with close contact among them). The IAV infection was investigated in OF by rRT-PCR testing and confirmed by viral isolation in cell culture. Results: We found 107 (7.4%) positives to IAV by rRT-PCR from a total of 1,444 OF samples tested. Additionally, 9 IAV isolates were all further identified as H1 subtype. Conclusions: Our results demonstrate that OF can be easily implemented as a novel, user-friendly, welfare-friendly, accurate and cost-effective sampling method for active surveillance and monitoring of IAV infections in swine farms in tropical settings.

Abstract Background The association of cough with Mycoplasma hyopneumoniae (MHP) DNA detection in specimens was evaluated under conditions in which the MHP status of inoculated and contact-infected pen mates was closely monitored for 59 days post-inoculation (DPI). Methods Seven-week-old pigs (n = 39) were allocated to five rooms (with one pen). Rooms contained 9 pigs each, with 1, 3, 6, or 9 MHP-inoculated pigs, respectively, except Room 5 (three sham-inoculated pigs). Cough data (2 × week) and specimens, tracheal swabs (2 × week), oral fluids (daily), drinker wipes (~ 1 × week), and air samples (3 × week) were collected. At 59 DPI, pigs were euthanized, and lung and trachea were evaluated for gross and microscopic lesions. Predictive cough value to MHP DNA detection in drinker and oral fluid samples were estimated using mixed logistic regression. Results Following inoculation, MHP DNA was first detected in tracheal swabs from inoculated pigs (DPI 3), then oral fluids (DPI 8), air samples (DPI 10), and drinker wipes (21 DPI). MHP DNA was detected in oral fluids in 17 of 59 (Room 1) to 43 of 59 (Room 3) samples, drinker wipes in 4 of 8 (Rooms 2 and 3) to 5 of 8 (Rooms 1 and 4) samples, and air samples in 5 of 26 (Room 2) or 3 of 26 (Room 4) samples. Logistic regression showed that the frequency of coughing pigs in a pen was associated with the probability of MHP DNA detection in oral fluids (P

Background: According to the World Health Organization, road traffic injuries lead to 1.3 million deaths each year and represent the leading cause of death for young adults under 30 years old. The use of psychoactive substances, including alcohol, drugs and pharmaceuticals, is a well-known risk factor for road traffic injuries. Our study aims to assess the prevalence of substances consumed by drivers in western Switzerland. Such studies are pivotal to improving prevention and developing public awareness campaigns. Methods: To assess the prevalence of psychoactive substances among drivers, roadside controls were performed in collaboration with local police, using their classical sampling procedures to detect drivers under the influence of drugs or alcohol over two time periods (P1: 2006-2008, P2: 2017-2020). When impaired driving was not suspected by the police, minimally invasive sampling strategies (i.e., oral fluids during P1 and dried blood spots during P2) were performed on volunteer drivers after a road safety survey. A posteriori analyses and statistical interpretation were then performed. Results: Among the 1605 drivers included in the study, 1048 volunteers provided an oral fluid sample, while 299 provided a dried blood spot sample. The percentage of drivers testing positive for at least one substance that can impact driving abilities was stable over time, with a rate of 10.5% positivity measured over both periods. Considering the different categories of substances, a slight variation was observed between both periods, with 7.6 and 6.3% of pharmaceuticals and 3.6 and 4.9% of illicit drugs for P1 and P2, respectively. Regarding the consumption of illicit drugs, the highest percentage of positivity was measured in biological fluids of drivers under the age of 35, during nights and week-ends, periods which are considered particularly prone to fatal accidents for this age group. Disturbingly, the road safety survey highlighted that drivers' perception of the risk of getting positively controlled while driving after drug consumption is low (3.3 on a 1-to-10 scale, N = 299). Conclusion: The number of positive cases measured in voluntary drivers who passed the preliminary police check demonstrates the importance of systematic biofluid sampling strategies regarding driving under the influence of psychoactive substances. Although the number of fatal road accidents globally has decreased over time, the results of this study reveal the need for both better prevention and deterrent processes that could potentially reduce the risk of fatal road accidents associated with drug consumption. [ABSTRACT FROM AUTHOR]

The use and abuse of cannabis, be it for medicinal or recreational purposes, is widely spread among the population. Consequently, a market for more potent and consequently more toxic synthetic cannabinoids has flourished, and with it, the need for accurate testing of these substances in intoxicated people. In this regard, one of the critical factors in forensic toxicology is the stability of these drugs in different biological matrices due to different storage conditions. This review aims to present the most updated and relevant literature of studies performed on the effects of different storage conditions on the stability of cannabis compounds present in various biological matrices, such as blood and plasma, urine, and oral fluids, as well as in alternative matrices, such as breath, bile fluid, hair, sweat, cerumen, and dried blood spots. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSV) has been one of the major health‐related concerns in the swine production industry. Through its rapid transmission and mutation, the simultaneous circulation of multiple PRRSV strains can be a challenge in PRRSV diagnostic, control and surveillance. The objective of this longitudinal study was to describe the temporal detection of PRRSV in swine farms with different production types and PRRS management strategies. Tonsil scraping (n = 344) samples were collected from three breeding and two growing herds for approximately one year. In addition, processing fluids (n = 216) were obtained from piglet processing batches within the three breeding farms while pen‐based oral fluids (n = 125) were collected in the two growing pig farms. Viral RNA extraction and reverse‐transcription quantitative PCR (RT‐qPCR) were conducted for all samples. The sample positivity threshold was set at quantification cycle (Cq) of ≤ 37. Statistical analyses were performed using generalized linear modelling and post hoc pairwise comparisons with Bonferroni adjustments using R statistical software. The results suggested a higher probability of detection in processing fluids compared to tonsil scraping specimens [odds ratio (OR) = 3.86; p =.096] in breeding farms whereas oral fluids were outperformed by tonsil scrapings (OR = 0.26; p <.01) in growing pig farms. The results described herein may lead to an improvement in PRRSV diagnostic and surveillance by selecting proper specimens. [ABSTRACT FROM AUTHOR]

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Objective: To assess the effects of oral fluids and intravenous fluids in terms of frequency of improving Amniotic Fluid Index during third trimester of pregnancy having oligohydramnios. Study Design: Comparative prospective survey. Place and Duration of Study: Obstetrics and Gynaecological Department of Fauji Foundation Hospital Rawalpindi, from Apr 2018 to Oct 2018. Methodology: A total of 100 patients with singleton pregnancy (50 in each group) at gestational age of 28-37 weeks and AFI of 5cm or less than 5cm were included. Non probability consecutive sampling technique was used. Ethics approval from hospital ethical committee and research board was taken. Patients were randomized into 2 groups (50 patients each) by lottery method. Group A was instructed to drink 2 liters of water per day for a time period of seven days and patients in group B was given 2 liters of 5% D/W which was in addition to their normal fluids intake. Amniotic fluid index was measured pre hydration and post hydration in both groups. Data was analyzed using SPSS-21. Results: Mean age of the patients was 33.62 ± 5.45 years and 34.70 ± 4.76 years for groups A and B respectively. In group A and group B mean gestational age was 34.28 ± 1.85 weeks vs 34.32 ± 1.82 weeks. In group A, 44 (88%)..............

Vertical transmission is a consistently discussed pathway of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3) transmission in pigs. To evaluate the presence of PCV2 and PCV3 in piglets, we collected tissue samples from 185 piglets that were crushed within the first week of life from 16 farms located in Germany and Austria. Pooled samples consisting of thymus, inguinal lymph node, myocardium, lung and spleen were examined for PCV2 and PCV3 by qPCR. Furthermore, oral fluid samples (OFS) from grow–finish pigs were collected and examined the same way. In piglets, PCV2 was highly prevalent (litters: 69.4%; piglets: 61.6%), whereas PCV3 prevalence was low (litters: 13.4%; piglets: 13.0%). In total, 72.6% and 67.2% of all collected OFS were PCV2 or PCV3 positive, respectively. Sow vaccination against PCV2 was identified as a protective factor concerning PCV2 in piglets (OR: 0.279; CI: 0.134–0.578; p < 0.001), whereas the porcine reproductive and respiratory syndrome virus (PRRSV) vaccination of sows was identified as a protective factor concerning PCV3 in piglets (OR: 0.252 CI: 0.104–0.610; p = 0.002). Our results show that PCV2, but not PCV3, is ubiquitous in suckling piglets and that early PCV3 infections might be modulated by PRRSV–PCV3 interaction. However, the ubiquitous nature of both viruses in older pigs could be confirmed. [ABSTRACT FROM AUTHOR]

Swine viral diseases challenge the sector's sustainability by affecting productivity and the health and welfare of the animals. The lack of antiviral drugs and/or effective vaccines renders early and reliable diagnosis the basis of viral disease management, underlining the importance of point-of-care (POC) diagnostics. A novel POC diagnostic device utilizing photonic integrated circuits (PICs), microfluidics, and information and communication technologies for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza A (SIV) was validated using spiked and clinical oral fluid samples. Metrics including sensitivity, specificity, accuracy, precision, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR) were calculated to assess the performance of the device. For PRRSV, the device achieved a sensitivity of 83.5%, specificity of 77.8%, and DOR values of 17.66, whereas the values for SIV were 81.8%, 82.2%, and 20.81, respectively. The POC device and PICs can be used for the detection of PRRSV and SIV in the field, paving the way for the introduction of novel technologies in the field of animal POC diagnostics to further optimize livestock biosecurity. [ABSTRACT FROM AUTHOR]

Abstract Recent decades have seen both rapid growth and extensive consolidation in swine production. As a collateral effect, these changes have exacerbated the circulation of viruses and challenged our ability to prevent, control, and/or eliminate impactful swine diseases. Recent pandemic events in human and animal health, e.g., SARS-CoV-2 and African swine fever virus, highlight the fact that clinical observations are too slow and inaccurate to form the basis for effective health management decisions: systematic processes that provide timely, reliable data are required. Oral fluid-based surveillance reflects the adaptation of conventional testing methods to an alternative diagnostic specimen. The routine use of oral fluids in commercial farms for PRRSV and PCV2 surveillance was first proposed in 2008 as an efficient and practical improvement on individual pig sampling. Subsequent research expanded on this initial report to include the detection of ≥23 swine viral pathogens and the implementation of oral fluid-based surveillance in large swine populations (> 12,000 pigs). Herein we compile the current information regarding oral fluid collection methods, testing, and surveillance applications in swine production.

Background: The association of cough with Mycoplasma hyopneumoniae (MHP) DNA detection in specimens was evaluated under conditions in which the MHP status of inoculated and contact-infected pen mates was closely monitored for 59 days post-inoculation (DPI). Methods: Seven-week-old pigs (n = 39) were allocated to five rooms (with one pen). Rooms contained 9 pigs each, with 1, 3, 6, or 9 MHP-inoculated pigs, respectively, except Room 5 (three sham-inoculated pigs). Cough data (2 × week) and specimens, tracheal swabs (2 × week), oral fluids (daily), drinker wipes (~ 1 × week), and air samples (3 × week) were collected. At 59 DPI, pigs were euthanized, and lung and trachea were evaluated for gross and microscopic lesions. Predictive cough value to MHP DNA detection in drinker and oral fluid samples were estimated using mixed logistic regression. Results: Following inoculation, MHP DNA was first detected in tracheal swabs from inoculated pigs (DPI 3), then oral fluids (DPI 8), air samples (DPI 10), and drinker wipes (21 DPI). MHP DNA was detected in oral fluids in 17 of 59 (Room 1) to 43 of 59 (Room 3) samples, drinker wipes in 4 of 8 (Rooms 2 and 3) to 5 of 8 (Rooms 1 and 4) samples, and air samples in 5 of 26 (Room 2) or 3 of 26 (Room 4) samples. Logistic regression showed that the frequency of coughing pigs in a pen was associated with the probability of MHP DNA detection in oral fluids (P < 0.01) and nearly associated with drinker wipes (P = 0.08). Pathology data revealed an association between the period when infection was first detected and the severity of gross lung lesions. Conclusions: Dry, non-productive coughs suggest the presence of MHP, but laboratory testing and MHP DNA detection is required for confirmation. Based on the data from this study, oral fluids and drinker wipes may provide a convenient alternative for MHP DNA detection at the pen level when cough is present. This information may help practitioners in specimen selection for MHP surveillance. [ABSTRACT FROM AUTHOR]

Abstract Salmonella is a leading cause of bacterial foodborne-related illness and pork products are a food-associated source. With > 50% of U.S. swine herds testing positive for Salmonella, asymptomatic carrier pigs that shed Salmonella in their feces are a food safety and environmental contamination issue. Herd level surveillance of Salmonella shedding status is useful, but collection of feces and culture methods for Salmonella detection are laborious and time-consuming. Surveillance for Salmonella-exposure through detection of Salmonella-specific serum antibody is a reliable method, but presents labor and animal-welfare issues. Oral fluids are a reliable, antemortem sample with proven utility for surveillance in the swine industry. We tested oral fluid samples as a potential non-invasive, repeatable sample type for the presence of Salmonella-specific antibodies. An indirect enzyme-linked immunosorbent assay (ELISA) detected anti-Salmonella IgG, IgM, and predominantly IgA in oral fluids from Salmonella enterica serovar Typhimurium-exposed pigs. Furthermore, with minor modifications, a commercial ELISA-based kit also detected Salmonella-specific antibodies in oral fluids. Collectively, oral fluids may serve as a prospective surveillance tool for herd level monitoring of Salmonella exposure.

Abstract Background Porcine circovirus 3 (PCV3) is an emerging etiological agent to the swine industry. However, its circulating status and genetic characteristics were still unclear in Henan, central China. Here, 318 porcine oral fluid specimens were collected from asymptomatic pigs in five farms and tested by PCR . Results The results showed that the positive rate of PCV3 was 12.3% (39/318) for the total samples, and 15.06% (25/166) in the stall-based samples, 9.21% (14/152) in the pen-based samples. Of the PCV3-positive samples, 41.0% were also positive for porcine circovirus 2 (PCV2). Nucleotide sequence comparison indicated that the 10 complete genomes and 34 capsid (cap) genes in this study shared 98.7–99.9% and 98–100% pairwise identities to each other, respectively. According to phylogenetic analysis and sequence alignment of cap gene, all the isolated sequences were clustered into 3 clades, including subgroup 1 (21/39, 61.8%), subgroup 2 (5/39, 14.7%) and subgroup 3 (8/39, 23.5%). Similar to previous reports, four amino acids (V24A, K27R, S77 T and I150L) in cap protein were identified as a conserved subgroup specific molecular marker. Conclusion Our research provided new insights into the epidemiology surveillance and genetic characteristics of PCV3 in China.

Active matrix metalloproteinase (aMMP)-8 utilized in point-of-care testing (POCT) is regarded as a potential biomarker for periodontal and peri-implant diseases. Various host and microbial factors eventually influence the expression, degranulation, levels and activation of aMMP-8. The type of oral fluids (saliva, mouthrinse, gingival crevicular, and peri-implant sulcular fluids [GCF/PISF], respectively) affect the analysis. With this background, we aimed to review here the recent studies on practical, inexpensive, noninvasive and quantitative mouthrinse and GCF/PISF chair-side POCT lateral flow aMMP-8 immunoassays (PerioSafe and ImplantSafe/ORALyzer) and how they help to detect, predict, monitor the course, treatment and prevention of periodontitis and peri-implantitis. The correlations of aMMP-8 POCT to other independent and catalytic activity assays of MMP-8 are also addressed. The mouthrinse aMMP-8 POCT can also detect prediabetes/diabetes and tissue destructive oral side-effects due to the head and neck cancers' radiotherapy. Chlorhexidine and doxycycline can inhibit collagenolytic human neutrophil and GCF aMMP-8. Furthermore, by a set of case-series we demonstrate the potential of mouthrinse aMMP-8 POCT to real-time/online detect periodontitis as a potential risk disease for coronavirus disease 2019 (COVID-19). The clinical interdisciplinary utilization of aMMP-8 POCT requires additional oral, medical, and interdisciplinary studies. [ABSTRACT FROM AUTHOR]

The use and abuse of cannabis, be it for medicinal or recreational purposes, is widely spread among the population. Consequently, a market for more potent and consequently more toxic synthetic cannabinoids has flourished, and with it, the need for accurate testing of these substances in intoxicated people. In this regard, one of the critical factors in forensic toxicology is the stability of these drugs in different biological matrices due to different storage conditions. This review aims to present the most updated and relevant literature of studies performed on the effects of different storage conditions on the stability of cannabis compounds present in various biological matrices, such as blood and plasma, urine, and oral fluids, as well as in alternative matrices, such as breath, bile fluid, hair, sweat, cerumen, and dried blood spots.

Background: Dengue is an acute viral infection and occurs in three different conditions- Dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome. Supportive and symptomatic treatment is indicated during infection, however less is known about feeding care obligatory in its management. Objective: it was aimed to study feeding care offered to DF patients admitted to hospitals. Materials and Method: An observational study was conducted on 48 patients (16 children and 32 adults) diagnosed for dengue fever and admitted to hospital (3 -government and 2-private hospitals were included) during the months of January to May. Details regarding medical symptoms, body temperature, state of consciousness, food tolerance and food intake from day 1 to day 5 was obtained. Oral intake of water and IV infused were recorded. Results: Relationship existed between symptoms, state of consciousness and tolerance to food and water; accordingly patients were classified as seriously sick, moderately sick and mildly sick. Patients with body temperature ˃102°F, abdomen pain, hepatomegaly and dehydration was associated with food intolerance and non-responsiveness (seriously sick), they took significantly longer time to develop food tolerance compared to their counterpart with less serious conditions (p ˂ 0.0001). Severity of sickness affected hospital stay, severely sick patients stayed longer as compared to the mildly sick (children 4.67±0.94 vs 4±0; adults 6.43±3.16 vs 3.67± 1.11 days). Majority of adults (71.9%) and children (56.3%) received normal diet. Energy and protein intake of patients (adults and children) in all the three groups were markedly lower than the RDA. Energy intake increased significantly from 1st to 5th day in all three groups. Differences in energy intake (adults-5.0±2.6 Kcals to 18.0±6.5 and children 10.0±3.8 - 29.0±6.8 Kcal/kg/day) was extremely significant (p˂ 0.001) among severely sick compared to the moderately (adults 11.0±1.6- 17.0±4.9 and children 20.0±2.9- 23.0±6.9 Kcal/kg/day) and mildly sick patients (adults 17.0±2.1- 21.0±3.8 and children 38.0±7.8- 39.0±5.7 Kcal/kg/day). Protein intake among both adults and children was very low. All patients regardless of the conditions received IV fluids at time of hospital admission. Severely sick received higher volumes of IV infusions and less quantity of oral fluids compared to the moderate and mildly sick. With improvement in conditions, it crisscrossed with reduced IV infusions and increased oral intakes. Each patient had different fluid needs and varied each day. Conclusion: Presently, diet management of DF patient is less judicious. In view of poor tolerance and unconsciousness that prevail, food selection should be carefully managed. Easily digestible and nutritious drinks such as fruits juices and soups should be included to complement for improving electrolyte disturbance and dehydration. Data base should be developed about feeding requirements for DF patients. [ABSTRACT FROM AUTHOR]

Vertical transmission is a consistently discussed pathway of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3) transmission in pigs. To evaluate the presence of PCV2 and PCV3 in piglets, we collected tissue samples from 185 piglets that were crushed within the first week of life from 16 farms located in Germany and Austria. Pooled samples consisting of thymus, inguinal lymph node, myocardium, lung and spleen were examined for PCV2 and PCV3 by qPCR. Furthermore, oral fluid samples (OFS) from grow–finish pigs were collected and examined the same way. In piglets, PCV2 was highly prevalent (litters: 69.4%; piglets: 61.6%), whereas PCV3 prevalence was low (litters: 13.4%; piglets: 13.0%). In total, 72.6% and 67.2% of all collected OFS were PCV2 or PCV3 positive, respectively. Sow vaccination against PCV2 was identified as a protective factor concerning PCV2 in piglets (OR: 0.279; CI: 0.134–0.578; p < 0.001), whereas the porcine reproductive and respiratory syndrome virus (PRRSV) vaccination of sows was identified as a protective factor concerning PCV3 in piglets (OR: 0.252 CI: 0.104–0.610; p = 0.002). Our results show that PCV2, but not PCV3, is ubiquitous in suckling piglets and that early PCV3 infections might be modulated by PRRSV–PCV3 interaction. However, the ubiquitous nature of both viruses in older pigs could be confirmed.

Objective: To assess the effects of oral fluids and intravenous fluids in terms of frequency of improving Amniotic Fluid Index during third trimester of pregnancy having oligohydramnios. Study Design: Comparative prospective survey. Place and Duration of Study: Obstetrics and Gynaecological Department of Fauji Foundation Hospital Rawalpindi, from Apr 2018 to Oct 2018. Methodology: A total of 100 patients with singleton pregnancy (50 in each group) at gestational age of 28-37 weeks and AFI of 5cm or less than 5cm were included. Non probability consecutive sampling technique was used. Ethics approval from hospital ethical committee and research board was taken. Patients were randomized into 2 groups (50 patients each) by lottery method. Group A was instructed to drink 2 liters of water per day for a time period of seven days and patients in group B was given 2 liters of 5% D/W which was in addition to their normal fluids intake. Amniotic fluid index was measured pre hydration and post hydration in both groups. Data was analyzed using SPSS-21. Results: Mean age of the patients was 33.62 ± 5.45 years and 34.70 ± 4.76 years for groups A and B respectively. In group A and group B mean gestational age was 34.28 ± 1.85 weeks vs 34.32 ± 1.82 weeks. In group A, 44 (88%) while in group B, 37 (74.0%) patients were of para 0-3 while para 4-7 were 6 (12.0%) in group A and 13 (26%) in group B. In group A 36 (72%) patients although in group B, 34 (68%) were of gravida 1-4. In group A and group B, 14 (28%) vs 16 (32%) were of gravida 5-8. Improvement was observed in 39 (78%) patients of group A while 22 (44%) patients of group B (p<0.001). In group A pre-hydration, mean amniotic fluid index level was 4.79 ± 0.53 and in group B 4.87 ± 0.36 (p=0.383). Post-hydration mean amniotic fluid index level was 6.79 ± 1.22 and 5.97 ± 1.37 in group A and B, respectively (p=0.002). Conclusion: Oral hydration of gestational women is more effective as compare to intravenous hydration in terms of frequency of improving Amniotic Fluid level during third trimester of pregnancy having oligohydramnios. [ABSTRACT FROM AUTHOR]

Background: Dental caries is an infectious and communicable disease and multiple factors influence the initiation and progression of the disease. Imbalance between oxidative stress and saliva antioxidants plays a major role in initiation and spread of dental caries. Objective: The aim of this study was to evaluate physicochemical properties of saliva such as flow rate, pH, buffering capacity, interleukin(IL)-6, IL-10, and malondialdehyde (MDA), and total antioxidants capacity (TAC), and catalase (CAT) levels in caries free and caries active individuals. Material and Method: Fifty consecutive subjects with dental caries experience (27 M & 23F) and fifty caries free subjects (24 M & 26 F) were recruited. The dental caries status was assessed depending on the WHO oral assessment form. Stimulated saliva samples were collected at the morning from both groups. The MDA, TAC, and CAT were evaluated using spectrophotometric assay. SPSS 13 analyzed data using Student's t-test. Results: There was a statistically significant increase in salivary T A C, CAT, and IL-6 in group with dental caries compared with control group. However, the salivary pH, flow rate, buffering capacity, and IL-10 levels were statistically lower in caries active patients. Person's correlation coefficient indicated significant negative correlations between the IL-6 with IL-10, pH, MDA, and flow rate. Whereas positive correlation with TAC, and buffering capacity. In addition, there was negative correlation between IL-10 with TAC, MDA, and buffering capacity, While positive correlation with pH and Flow rate. Conclusions: Alterations in salivary MDA, TAC, CAT, and IL-6 levels were significantly higher whereas Salivary pH, flow rate, buffering capacity, and IL-10 were significantly lower in the study groups when compared to control groups. [ABSTRACT FROM AUTHOR]

Aerosols are suspensions of liquid and solid particles in the atmosphere, excluding clouds and precipitation. The aerosol particle sizes range from 10-4 to 10 pm. The purpose of this study was to focus attention on the need to adopt infection control procedures in dentistry. The aerosol and microbial load in various dental clinics during routine procedures were determined. Oral fluids become aerosolized during dentistry and oral microbes have been used as the markers of their spread that may carry blood-borne pathogens. In dental clinics, aerosols are produced while using dental handpieces, such as ultrasonic scalers, air rotors, micromotors and/or air-water syringes. These bio-aerosols consist of droplets, also known as spatters, and droplet nuclei. Droplets, sized >100 pm, settle rapidly onto surfaces in the immediate proximity of the source. Droplet nuclei, <10 pm, are lighter and, therefore, can remain in the air for hours before settling on a surface. These aerosols contain microorganisms originating from the patient's oral cavity and from dental unit waterlines (DUWLs). The release of microorganisms into aerosols increases the microbial burden in the air and can lead to the contamination of all surfaces in a dental treatment room. Because of the frequent aerosol generating procedures in dental practice, these aerosols can function as an important mode for infection transmission in dental clinics. [ABSTRACT FROM AUTHOR]

Madin–Darby canine kidney (MDCK) cells are commonly used for the isolation of mammalian influenza A viruses. The goal of this study was to compare the sensitivity and suitability of the original MDCK cell line in comparison with MDCK-derived cell lines, MDCK.2, MDCK SIAT-1 and MDCK-London for isolation of swine-origin influenza A viruses (IAV-S) from clinical specimens. One-hundred thirty clinical specimens collected from pigs in the form of nasal swabs, lung tissue and oral fluids that were positive by PCR for the presence of IAV-S RNA were inoculated in the cell cultures listed above. MDCK-SIAT1 cells yielded the highest proportion of positive IAV-S isolations from all specimen types. For nasal swabs, 58.62% of the specimens were IAV-S positive in MDCK-SIAT1 cells, followed by MDCK-London (36.21%), and conventional MDCK and MDCK.2 cells (27.5%). For lung specimens, 59.38% were IAV-S positive in MDCK-SIAT1 cells, followed by MDCK-London (40.63%), and conventional MDCK and MDCK.2 cells (18.75–31.25%). Oral fluids yielded the lowest number of positive virus isolation results, but MDCK-SIAT1 cells were still had the highest rate (35%) of IAV-S isolation, whereas the isolation rate in other cells ranged from 5–7.5%. Samples with lower IAV-S PCR cycle threshold (Ct) values were more suitable for culturing and isolation. The isolated IAV-S represented H1N1-β, H1N2-α, H1N1pdm and H3N2 cluster IV and cluster IVB viruses. The result of the current study demonstrated the importance of using the most appropriate MDCK cells when isolating IAV-S from clinical samples.

Abstract Background The usefulness of oral fluid (OF) sampling for surveillance of infections in pig populations is already accepted but its value as a tool to support investigations of porcine respiratory disease complex (PRDC) has been less well studied. This study set out to describe detection patterns of porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), swine influenza virus type A (SIV) and Mycoplasma hyopneumoniae (M. hyo) among farms showing differing severity of PRDC. The study included six wean-to-finish pig batches from farms with historical occurrence of respiratory disease. OF samples were collected from six pens every two weeks from the 5th to the 21st week of age and tested by real time PCR for presence of PRRSV, SIV and M. hyo and by quantitative real time PCR for PCV2. Data was evaluated alongside clinical and post-mortem observations, mortality rate, slaughter pathology, histopathology, and immunohistochemistry testing data for PCV2 antigen where available. Results PRRSV and M. hyo were detectable in OF but with inconsistency between pens at the same sampling time and within pens over sequential sampling times. Detection of SIV in clinical and subclinical cases showed good consistency between pens at the same sampling time point with detection possible for periods of 2–4 weeks. Quantitative testing of OF for PCV2 indicated different patterns and levels of detection between farms unaffected or affected by porcine circovirus diseases (PCVD). There was good correlation of PCR results for multiple samples collected from the same pen but no associations were found between prevalence of positive test results and pen location in the building or sex of pigs. Conclusions Detection patterns for PRRSV, SIV and M. hyo supported the effectiveness of OF testing as an additional tool for diagnostic investigation of PRDC but emphasised the importance of sampling from multiple pens and on multiple occasions. Preliminary evidence supported the measurement of PCV2 load in pooled OF as a tool for prediction of clinical or subclinical PCVD at farm level.

Background and objectives: Childbirth consists of a three-stage period beginning with uterine contractions and ending in the expulsion of the placenta. The present study was conducted to compare the effects of Ringer's solution, dextrose solution 5%, and oral intake on the duration of total labor and labor stages (primary outcome) and the frequency of oxytocin administration and prolonged labor (secondary outcome) in nulliparous women with spontaneous vaginal delivery.Materials and methods: The present randomized clinical trial was conducted on 201 nulliparous women in labor assigned with a randomized block design to three groups, each receiving either Ringer's solution plus oral fluids, dextrose solution 5% solution plus oral fluids or oral fluids alone. The solutions were intravenously administered at a rate of 125 ml/h in the groups receiving the solutions and the subjects in the oral fluids group could consume liquids of their choice (water, orange juice, and apple juice). The duration of total labor and labor stages (the active stage, the second, and third stages of labor) were recorded by the researcher in minutes. Furthermore, the frequency of oxytocin administration and prolonged labor were recorded by the researcher by using a checklist. Data were analyzed using the Kruskal-Wallis test, Mann-Whitney's U-test, chi- square test and Fisher exact test.Results: The mean (±SD) duration of total labor was 301.2 (±50.0) minutes in the Ringer group, 171.9 (±36.4) in the dextrose 5% group, and 524.8 (±103.4) in the oral fluids group, suggesting a statistically significant intergroup difference (p < .001). The mean (±SD) duration of the active stage of labor was 276.7 (±91.3) minutes in the Ringer group, 150.6 (±78.5) in the dextrose 5% group, and 302.4 (±136.8) in the oral fluids group, suggesting a statistically significant intergroup difference (p < .001). A significant difference was also observed between the three groups in terms of the duration of the third stage of labor (p < .001); however, no significant differences were observed in terms of the duration of the second stage of labor (p = .058). Furthermore, a significant difference was observed between the three groups in terms of the need for oxytocin administration (p ≤ .001) and the frequency of prolonged labor (p = .001).Conclusion: The results suggest that the consumption of dextrose solution by nulliparous women reduces the duration of labor, the need for oxytocin administration and the frequency of prolonged labor compared to when Ringer's solution and oral fluids are used. [ABSTRACT FROM AUTHOR]

Salmonella is a leading cause of bacterial foodborne-related illness and pork products are a food-associated source. With > 50% of U.S. swine herds testing positive for Salmonella, asymptomatic carrier pigs that shed Salmonella in their feces are a food safety and environmental contamination issue. Herd level surveillance of Salmonella shedding status is useful, but collection of feces and culture methods for Salmonella detection are laborious and time-consuming. Surveillance for Salmonella-exposure through detection of Salmonella-specific serum antibody is a reliable method, but presents labor and animal-welfare issues. Oral fluids are a reliable, antemortem sample with proven utility for surveillance in the swine industry. We tested oral fluid samples as a potential non-invasive, repeatable sample type for the presence of Salmonella-specific antibodies. An indirect enzyme-linked immunosorbent assay (ELISA) detected anti-Salmonella IgG, IgM, and predominantly IgA in oral fluids from Salmonella enterica serovar Typhimurium-exposed pigs. Furthermore, with minor modifications, a commercial ELISA-based kit also detected Salmonella-specific antibodies in oral fluids. Collectively, oral fluids may serve as a prospective surveillance tool for herd level monitoring of Salmonella exposure. [ABSTRACT FROM AUTHOR]

Marijuana (i.e. , cannabis) and its derivatives are considered the most commonly used of illicit drugs. Within the last two decades, phytocannabinoids and their synthetic analogues have emerged as potential medicines for the treatment of various disorders via targeting of the endocannabinoid system. Recently, some countries have legalized (medicinal/recreational) cannabis, which now allows for more research to be conducted. Accordingly, sensitive and specific analytical assays are required to identify and quantify these compounds in different human matrices. These analytical approaches were developed using mass spectrometric detection, where LC–MS/MS specifically has become the mainstay for the quantitative analysis of tetrahydrocannabinol and other cannabinoids. This is due to its superior selectivity and sensitivity, and ability to determine free and conjugate analytes within the same analysis. This tabular review of such methods is prefaced by a short overview of the major cannabinoids and some of their physiological actions. Image 1 • Literature review 2012 to 2018. • LC-MS detect and quantify cannabinoids beyond ppb level in various biological matrices. • LC methods are preferable compared to GC techniques. • LC methods do not require time-consuming or complicated sample preparations procedures and can facilitate automation of cannabinoids analysis. [ABSTRACT FROM AUTHOR]

Mycoplasma hyorhinis (Mhr) and M. hyosynoviae (Mhs) are commensal organisms of the upper respiratory tract and tonsils but may also cause arthritis in pigs. In this study, 8-week-old cesarean-derived colostrum-deprived (CDCD) pigs (n = 30; 3 groups, 10 pigs per group, 2 pigs per pen) were inoculated with Mhr , Mhs , or mock-inoculated with culture medium and then pen-based oral fluids were collected at different time points over the 56 days of the experimental study. Oral fluids tested by Mhr and Mhs quantitative real-time PCRs revealed Mhr DNA between day post inoculation (DPI) 5–52 and Mhs DNA between DPI 5–15. Oral fluids were likewise tested for antibody using isotype-specific (IgG, IgA, IgM) indirect ELISAs based on a recombinant chimeric polypeptide of variable lipoproteins (A-G) for Mhr and Tween 20-extracted surface proteins for Mhs. Mhr IgA was detected at DPI 7 and, relative to the control group, significant (p < 0.05) antibody responses were detected in the Mhr group between DPI 12–15 for IgM and DPI 36–56 for both IgA and IgG. In the Mhs group, IgM was detected at DPI 10 and significant (p < 0.05) IgG and IgA responses were detected at DPI 32–56 and DPI 44–56, respectively. This study demonstrated that oral fluid could serve as an effective and convenient antemortem sample for monitoring Mhr and Mhs in swine populations. • Mycoplasma hyorhinis and Mycoplasma hyosynoviae have emerged as important contributors of arthritis and lameness in growing pigs. • The infection dynamics of M. hyorhinis and M. hyosynoviae was evaluated in relation to the antibody response and bacterial shedding in oral fluids as antemortem specimen. • Oral fluid could serve as an effective and convenient antemortem sample for monitoring M. hyorhinis and M. hyosynoviae in swine populations. [ABSTRACT FROM AUTHOR]

Introduction: Diabetes mellitus has been linked with an increased risk of caries, gingivitis and periodontal disease. Dental caries is more prevalent and even more severe in diabetic patients than non-diabetics. The aetiology and pathogenesis of dental caries are known to be multifactorial. The secretion rates and quality of saliva are important not only in caries development but also for remineralisation. As there is an alteration of the salivary constituents in diabetic patients its constituents has gained much importance as a diagnostic and therapeutic tool. Various factors that regulate the caries activity balance are the pH of saliva and concentration of various ions including free calcium, phosphate, sodium, chloride, potassium and fluoride ions in the saliva. Aim: To investigate the association of salivary electrolyte concentration with dental caries among diabetic and nondiabetic individuals, of different age groups. Materials and Methods: Ninety-six Patients were evaluated for the study in which forty eight were considered for control and forty eight for diabetic, in which these control group and diabetic group were further subdivided into young adults (20-39 yrs) and old age adults (40-64 yrs) with twenty four patients each. Young adult age group and adult age group had sub groups with caries and without caries group having twelve patients each respectively. Saliva was collected from the individuals, centrifuged and the supernatant obtained was assessed for the salivary electrolytes: sodium, potassium, calcium, phosphorous and chloride levels using an Erba autoanalyser. The results obtained were tabulated, statistically analysed using Independent sample t-test and conclusions drawn. Results: The salivary electrolytes– sodium, potassium, calcium, and phosphorous showed a significant decrease in caries active diabetic patients of young adult group. But in the older adult group K and Cl levels were statistically higher in caries active patients. Potassium & Phosphorous were statistically higher in young adult age group (Diabetic and Non diabetic) without caries and Potassium only in adult age group. Conclusion: Salivary electrolyte plays a significant role in prevalence of dental caries in young diabetic individuals when compared to non-diabetic individuals and old age adults with and without diabetics. Therefore, maintaining the salivary electrolyte concentration in young diabetic individuals helps in remineralisation of the tooth which prevents decay.

ABSTRACT Human papillomavirus (HPV) vaccination elicits high-titer genotype-specific antibody responses that are associated with a reduced risk of cervical disease caused by vaccine-incorporated genotypes. Our objective was to evaluate dried blood spots (DBSs) and oral mucosal transudate (OMT) as alternative samples to serum to confirm HPV vaccine antibody status. A study was carried out to evaluate the feasibility of detecting HPV16 and HPV18 antibodies in OMT, DBSs, and sera among women who self-reported being unvaccinated or fully vaccinated with the HPV vaccine. Serum had the highest sensitivity (100%) for detection of antibodies against both HPV16 and HPV18 but the lowest specificity, due to the detection of natural infection antibodies in 16% of unvaccinated women. Conversely, DBSs and OMT had lower sensitivity (96% and 82%, respectively) but high specificity (98%). We confirmed that these antibodies were functional (i.e., neutralizing) and that their detection was quantitatively reproducible and well correlated between sample types when normalized to IgG content. DBSs and OMT are appropriate alternative sample types for HPV vaccine surveillance. These alternative sample types warrant consideration for the purposes of cervical screening, diagnosis, and management, but more work will be needed to establish the stringent parameters required for such application. IMPORTANCE Human papillomavirus (HPV) is the causative agent of cervical and other anogenital cancers. HPV vaccination, primarily targeted at young girls before the age of sexual debut, is starting to demonstrate population-level declines in HPV infection and early disease associated with vaccine-incorporated genotypes. Monitoring young women for vaccine-specific antibody is important for vaccine surveillance and may be useful as an adjunct test within a cervical screening context. We evaluated serum, dried blood spots, and oral fluid as potential samples for such applications and report robust measures of diagnostic accuracy. This is the first time a direct comparison of alternative sample types has been made between vaccinated and unvaccinated women for the detection and quantitation of HPV antibodies.

Introduction: Diabetes mellitus has been linked with an increased risk of caries, gingivitis and periodontal disease. Dental caries is more prevalent and even more severe in diabetic patients than non-diabetics. The aetiology and pathogenesis of dental caries are known to be multifactorial. The secretion rates and quality of saliva are important not only in caries development but also for remineralisation. As there is an alteration of the salivary constituents in diabetic patients its constituents has gained much importance as a diagnostic and therapeutic tool. Various factors that regulate the caries activity balance are the pH of saliva and concentration of various ions including free calcium, phosphate, sodium, chloride, potassium and fluoride ions in the saliva. Aim: To investigate the association of salivary electrolyte concentration with dental caries among diabetic and non-diabetic individuals, of different age groups. Materials and Methods: Ninety-six Patients were evaluated for the study in which forty eight were considered for control and forty eight for diabetic, in which these control group and diabetic group were further subdivided into young adults (20-39 yrs) and old age adults (40-64 yrs) with twenty four patients each. Young adult age group and adult age group had sub groups with caries and without caries group having twelve patients each respectively. Saliva was collected from the individuals, centrifuged and the supernatant obtained was assessed for the salivary electrolytes: sodium, potassium, calcium, phosphorous and chloride levels using an Erba autoanalyser. The results obtained were tabulated, statistically analysed using Independent sample t-test and conclusions drawn. Results: The salivary electrolytes– sodium, potassium, calcium, and phosphorous showed a significant decrease in caries active diabetic patients of young adult group. But in the older adult group K and Cl levels were statistically higher in caries active patients. Potassium & Phosphorous were statistically higher in young adult age group (Diabetic and Non diabetic) without caries and Potassium only in adult age group. Conclusion: Salivary electrolyte plays a significant role in prevalence of dental caries in young diabetic individuals when compared to non-diabetic individuals and old age adults with and without diabetics. Therefore, maintaining the salivary electrolyte concentration in young diabetic individuals helps in remineralisation of the tooth which prevents decay. [ABSTRACT FROM AUTHOR]