Groupwise antibiotic treatments are common in broiler chicken production. They induce selection for antibiotic resistance in commensalEscherichia coli.This study aimed to investigate antibiotic resistance after individual (I, drenching) or groupwise treatment (G, by water) with amoxicillin, and after contact with I or G (KI or KG), compared with untreated broilers without contact with treated broilers (C), and pretreatment values. Finally, we compared antibiotic resistance from broilers (G) after a second treatment, with a treatment in the contact animals (KG), and a first treatment in the control animals (C). Resistance to ampicillin and other antibiotics was significantly increased in groups G and I within 2 days, suggesting (co-)selection of resistance. The increase was lower in groups KI, KG, and C during the first treatment (days 1-5). The increased resistance in group C was interpreted as a change in the microbiota after initial moving and first feeding. After treatment, resistance rates decreased to initial or lower values in all groups. During the second treatment period (days 34-38), all three groups' (G, KG, and C) resistance levels increased to equally high levels. Cephalosporin resistance was low, and did not change over the experimental period. On days 3 and 38, resistance rates ofE. colifrom duodenum, jejunum, and cecum did not differ between segments and treatment routes. Overall, the baseline levels of antibiotic resistance inE. coliwere high. Amoxicillin triggered an increase in resistance levels, irrespective of the mode of treatment. Substantial resistance dynamics in untreated controls warrant further investigation.

Brucellosis is reportedly endemic in ruminants in Pakistan. BothBrucella abortusandB. melitensisinfections have been decumented in domestic animals and humans in the country. This study aimed to identify the burden of anti-Brucellaantibodies in small ruminants as well as associated potential risk factors with its occurrence at nine institutional livestock farms in Punjab, Pakistan. The sera collected from equal number of sheep and goats (500 from each species) were screened by indirect-ELISA for anti-smooth-Brucellaantibodies followed by a serial detection by real-time PCR. Overall, 5.1% (51/1000) seropositivity was registered corresponding to 5% (25/500) prevalence in goats and 5.2% (26/500) in sheep.Brucella-DNA could not be detected in any of the tested sera by real-time PCR. Multiple logistic regression model indicated that farm location (OR 34.05), >4 years of age (OR 2.88), with history of reproductive disorders (OR 2.69), and with BCS of <= 3 (OR 12.37) were more likely to test positive for brucellosis at these farms. A routine screening, stringent biosecurity, and quarantine measures are warranted for monitoring and eradication of the infection. Similarly, isolation and molecular investigation of the etiologic agent(s) are needed to understand the relationship of epidemiology and out-breaks of brucellosis in the country.

Objectives: Navigated transcranial magnetic stimulation (nTMS) provides significant benefits over classic TMS. Yet, the acquisition of individual structural magnetic resonance images (MRIindividual) is a time-consuming, expensive, and not feasible prerequisite in all subjects for spatial tracking and anatomical guidance in nTMS studies. We hypothesize that spatial transformation can be used to adjust MRI templates to individual head shapes (MRIwarped) and that TMS parameters do not differ between nTMS using MRIindividual or MRIwarped.

Materials and Methods: Twenty identical TMS sessions, each including four different navigation conditions, were conducted in 10 healthy subjects (one female, 27.4 ± 3.8 years), i.e., twice per subject by two researchers to additionally assess interrater reliabilities. MRIindividual were acquired for all subjects. MRIwarped were obtained through the spatial transformation of a template MRI following a 5-, 9-and 36-point head surface registration (MRIwarped_5, MRIwarped_9, MRIwarped_36). Stimulation hotspot locations, resting motor threshold (RMT), 500 μV motor threshold (500 μV-MT), and mean absolute motor evoked potential difference (MAD) of primary motor cortex (M1) examinations were compared between nTMS using either MRIwarped variants or MRIindividual and non-navigated TMS.

Results: M1 hotspots were spatially consistent between MRIindividual and MRIwarped_36 (insignificant deviation by 4.79 ± 2.62 mm). MEP thresholds and variance were also equivalent between MRIindividual and MRIwarped_36 with mean differences of RMT by -0.05 ± 2.28% maximum stimulator output (%MSO; t (19) = -0.09, p = 0.923), 500 μV-MT by -0.15 ± 1.63%MSO (t (19) = -0.41, p = 0.686) and MAD by 70.5 ± 214.38 μV (t (19) = 1.47, p = 0.158). Intraclass correlations (ICC) of motor thresholds were between 0.88 and 0.97.

Conclusions: NTMS examinations of M1 yield equivalent topographical and functional results using MRIindividual and MRIwarped if a sufficient number of registration points are used.

The reliability of transferring species distribution models (SDMs) to new ranges and future climates has been widely debated. Biological invasions offer the unique opportunity to evaluate model transferability, as distribution data between species' native and introduced ranges are geographically independent of each other. Here, we performed the first global quantitative synthesis of the spatial transferability of SDMs for 235 invasive species and assessed the association of model transferability with the focal invader, model choice and parameterisation. We found that SDMs had limited spatial transferability overall. However, model transferability was higher for terrestrial endotherms, species introduced from or to the Southern Hemisphere, and species introduced more recently. Model transferability was also positively associated with the number of presences for model calibration and evaluation, respectively, but negatively with the number of predictors. These findings highlight the importance of considering the characteristics of the focal invader, environment and modelling in the application and assessment of SDMs.

The Rum Layered Suite, NW Scotland, hosts Cr-spinel seams at the bases of peridotite-troctolite macro-rhythmic units in the eastern portion of the intrusion. Here, we present detailed field observations together with microstructural and mineral chemical analyses for the Unit 7-8 Cr-spinel seam and associated cumulates in the Eastern Layered Intrusion. Detailed mapping and sampling reveal significant lateral variations in the structural characteristics and mineral compositions of the Unit 7-8 boundary zone rocks. Although the Cr-spinel seam is laterally continuous over similar to 3 km, it is absent towards the centre and the margins of the intrusion. The compositional characteristics of Cr-spinel and plagioclase vary systematically along strike, exhibiting a chemical evolution towards more differentiated compositions with increasing distance from the main feeder conduit of the Rum intrusion; the Long Loch Fault. On the basis of our combined datasets, we propose that the upper part of the troctolite, the anorthosite layer underlying the Cr-spinel seam and the seam itself formed during a multi-stage magma replenishment event. The stages can be summarised as follows: (1) peridotite schlieren and anorthosite autoliths formed following melt infiltration and cumulate assimilation in the crystal mush of the Unit 7 troctolite. (2) The anorthosite layer then formed from the Unit 7 troctolite crystal mush by thermal erosion and dissolution due to infiltrating magma. (3) Subsequent dissolution of the anorthosite layer by new replenishing magma led to peritectic in situ crystallisation of the Unit 7-8 Cr-spinel seam, with (4) continued magma input eventually producing the overlying Unit 8 peridotite. In the central part of the Rum Layered Suite, the aforementioned assimilation of the troctolitic footwall formed the anorthosite layer. However, the absence of anorthosite in close proximity to the Long Loch Fault can be explained by enhanced thermochemical erosion close to the feeder zone, and its absence close to the margins of the intrusion, at maximum distance from the Long Loch Fault, may be due to cooling of the magma and loss of erosion potential. In line with other recent studies on PGE-bearing chromitites in layered intrusions, we highlight the importance of multi-stage intrusive magma replenishment to the formation of spatially coupled anorthosite and Cr-spinel seams, as well as the lateral mineral chemical variations observed in the Unit 7-8 boundary zone cumulates.

Background: Sleepiness at the wheel affects 10% to 15% of drivers and is one major cause of death on highways with one-third of fatal accidents. Obstructive sleep apnea (OSA) is one of the most common sleep disorders leading to sleepiness at the wheel. The aim of this study was to compare the psychomotor vigilance test reaction time (PVT RT) in OSA patients and controls (morning and afternoon) with the results of a divided attention steering simulator (DASS). A second purpose was to compare these results with the mean sleep latencies in the multiple sleep latency test (MSLT), the Epworth Sleepiness Scale (ESS) values and a neurocognitive test (test of attentional performance, TAP).

Patients and methods: Thirty eight OSA patients and 16 age and sex matched healthy controls were investigated by ESS, PVT, TAP, MSLT, and DASS (response time, failed responses, lane deviation, and off-road-events).

Results: With increasing age, the performance in the DASS decreased. There was no correlation between the DASS and the results of the MSLT and ESS. The controls showed a significantly faster DASS response time in the morning compared to OSA patients (median 2.1 versus 3.0; p=0.044) and fewer off-road events (9 versus 37; p=0.042). We found a moderate correlation between the PVT RT and all parameters of the DASS, as well as the TAP "alertness" subtest.

Conclusion: The increase of PVT RT as well as the decreased tonic alertness in the TAP in untreated OSA patients correlated with an impairment of simulated driving performance. The PVT and the TAP are both suitable diagnostic tools for measuring impaired driving ability in OSA patients. The MSLT did not correlate with the simulated driving performance. We recommend investigation of a longer version of the PVT in order to increase its sensitivity.

Laboratory male mice are often housed individually due to aggressive behavior or experimental requirements, though social isolation can cause welfare issues. As a strategy to refine housing of male mice, we introduce the separated pair housing system. A perforated transparent wall divides the cage into two compartments and allows olfactory, acoustic, and visual communication between the two mice but prevents fighting and injuries. Long-term effects of separated pair housing on well-being and distress of adult male C57BL/6JRj mice were investigated and compared with both single- and group-housed mice. Behavioral analysis after eight weeks in three different housing systems revealed no differences in burrowing performance, social interaction, anxiety, and stress hormone concentrations. However, pair-housed mice built more complex nests compared to single-housed mice and the nest position suggested that pair-housed mice preferred the close proximity to their cage mates. Moreover, pair-housed mice showed less locomotor activity compared to group- and single-housed mice. Body weight was higher in group-housed mice. All in all, no unambiguous long-term beneficial effects of pair housing on the well-being were found. However, the findings emphasized that effects of the housing systems on behavioral, physical, and biochemical parameters must be considered in the design of animal experimental studies.

Objective: Obsessive-compulsive disorder (OCD) is a complex psychiatric disorder with a substantial genetic contribution. While the specific variants underlying OCD's heritability are still unknown, findings from genome-wide association studies (GWAS) corroborate the importance of common SNPs explaining the phenotypic variance in OCD. Investigating associations between the genetic liability for OCD, as reflected by a polygenic risk score (PRS), and potential endophenotypes of the disorder, such as the personality trait harm avoidance, may aid the understanding of functional pathways from genes to diagnostic phenotypes. Methods: We derived PRS for OCD at severalP-value thresholds based on the latest Psychiatric Genomics Consortium OCD GWAS (2688 cases, 7037 controls) in an independent sample of OCD patients (n = 180), their unaffected first-degree relatives (n = 108) and healthy controls (n = 200). Using linear regression, we tested whether these PRS are associated with the personality trait harm avoidance. Results: Results showed that OCD PRS significantly predicted OCD status, with patients having the highest scores and relatives having intermediate scores. Furthermore, the genetic risk for OCD was associated with harm avoidance across the entire sample, and among OCD patients. As indicated by mediation analyses, harm avoidance mediated the association between the OCD PRS and OCD caseness. These results were observed at multipleP-value thresholds and persisted after the exclusion of patients with a current comorbid major depressive or anxiety disorder. Conclusion: Our findings support the polygenic nature of OCD and further validate harm avoidance as a candidate endophenotype and diathesis of OCD.

The aim of this study is to develop nanometer-thin epoxy-based films on aluminium alloy AA2024-T3 as a model coating system for high resolution corrosion studies. Spin coating was used for the layer-by-layer (LbL) deposition of poly-(ethylenimine) (PEI) and poly([o-cresyl glycidyl ether]-co-formaldehyde) (CNER) bilayers. The film chemistry and the cross-linking process were characterized by means of Fourier-transform infrared spectroscopy (FTIR). Ellipsometric data confirmed the linear increase of film thickness. The potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) results indicate the improvement of the film barrier properties with increasing film thickness. Mapping of the topography and the volta potential was performed by means of scanning Kelvin probe force microscopy (SKPFM). The results indicate the presence of a homogeneous film structure, while the intermetallic phases can still be identified below the coating. The SKPFM analysis confirmed that the model films are suitable for investigation of corrosion processes at the coating/metal interface.

Isolation of biomolecules in vacuum facilitates characterization of the intramolecular interactions that determine three-dimensional structure, but experimental quantification of conformer thermochemistry remains challenging. Infrared spectroscopy of molecules trapped in helium nanodroplets is a promising methodology for the measurement of thermochemical parameters. When molecules are captured in a helium nanodroplet, the rate of cooling to an equilibrium temperature ofca.0.4 K is generally faster than the rate of isomerization, resulting in "shock-freezing" that kinetically traps molecules in local conformational minima. This unique property enables the study of temperature-dependent conformational equilibriaviainfrared spectroscopy at 0.4 K, thereby avoiding the deleterious effects of spectral broadening at higher temperatures. Herein, we demonstrate the first application of this approach to ionic species by coupling electrospray ionization mass spectrometry (ESI-MS) with helium nanodroplet infrared action spectroscopy to probe the structure and thermochemistry of deprotonated DNA dinucleotides. Dinucleotide anions were generated by ESI, confined in an ion trap at temperatures between 90 and 350 K, and entrained in traversing helium nanodroplets. The infrared action spectra of the entrained ions show a strong dependence on pre-pickup ion temperature, consistent with the preservation of conformer population upon cooling to 0.4 K. Non-negative matrix factorization was utilized to identify component conformer infrared spectra and determine temperature-dependent conformer populations. Relative enthalpies and entropies of conformers were subsequently obtained from a van't Hoff analysis. IR spectra and conformer thermochemistry are compared to results from ion mobility spectrometry (IMS) and electronic structure methods. The implementation of ESI-MS as a source of dopant molecules expands the diversity of molecules accessible for thermochemical measurements, enabling the study of larger, non-volatile species.

Inhibitory control, the inhibition of prepotent actions, is essential for higher-order cognitive processes such as planning, reasoning, and self-regulation. Individuals and species differ in inhibitory control. Identifying what influences inhibitory control ability within and between species is key to understanding how it evolved. We compared performance in the cylinder task across five lizard species: tree skinks (Egernia striolata), gidgee skinks (Egernia stokesii), eastern blue-tongue skinks (Tiliqua s. scincoides), sleepy lizards (Tiliqua r. asper), and eastern water skinks (Eulamprus quoyii). In our task, animals had to inhibit the prepotent motor response of directly approaching a reward placed within a semi-transparent mesh cylinder and instead reach in through the side openings. Additionally, in three lizard species, we compared performance in the cylinder task to reversal learning to determine the task specificity of inhibitory ability. Within species, neither sex, origin, body condition, neophobia, nor pre-experience with other cognitive tests affected individual performance. Species differed in motor response inhibition: Blue-tongue skinks made fewer contacts with the semi-transparent cylinder wall than all other species. Blue-tongue skinks also had lower body condition than the other species which suggest motivation as the underlying cause for species differences in task performance. Moreover, we found no correlation between inhibitory ability across different experiments. This is the first study comparing cylinder task performance among lizard species. Given that inhibitory control is probably widespread in lizards, motor response inhibition as exercised in the cylinder task appears to have a long evolutionary history and is likely fundamental to survival and fitness. Significance The study of lizard cognition is receiving increasing attention. Lizards are a diverse group with a wide range of ecological attributes and represent a model system through which we can test a wide range of hypotheses relating to cognitive evolution. Furthermore, considering their evolutionary history, studying non-avian reptile cognition can help understand the evolution of different cognitive abilities including inhibitory control. Here, we provide a comparison of inhibitory control ability in five lizard species. Consequently, we are able to, firstly, validate a method (the cylinder task) initially developed for the use in mammals and birds, for use in lizards, and secondly, collect valuable data on inhibitory control in a poorly studied group with respect to cognitive ability. Our study suggests non-cognitive factors as a major influence on cylinder task performance, which is in agreement with previous studies of other vertebrates.

Background Blood profile testing is commonly used to monitor herd health status, diagnose disorders, and predict the risk of diseases in cows and calves, with subsequent optimization the production of dairy herds. By understanding the physiological ranges of serum metabolites relative to age, lactation stage, and the sampling time in healthy cows and calves, the dairy practitioners can accurately diagnose abnormalities with a blood test. The effect of sampling time on the variation of serum metabolites within 24 h were evaluated in 83 cattle. All animals were originated from a dairy herd, where the animals, based on their ages and lactation stages, were classified into eight groups. The blood samples were collected from each animal every 4 h within a day. Results The time of sampling within the day showed significant influences on the serum concentrations of glucose, beta-hydroxybutyric acid (BHBA) and urea. BHBA was the most metabolite that showed day variation among cows' groups. Furthermore, the concentrations of total cholesterol were the most stable metabolite in all groups. The mean values of albumin, total proteins, glucose, non-esterified fatty acids (NEFA), BHBA, total cholesterol, total bilirubin, urea, and creatinine revealed significant variations among the different studied groups. Conclusions A certain suitable time of blood sample collection cannot be recommended. However, care shall be taken for the time of sampling for measurements of glucose, NEFA, BHBA and urea, otherwise the comparative values of these metabolites at different sampling time points may differ significantly from each other's, without a disease cause. It may be recommended, for metabolic assessment of dairy herds, classification the subjects into different groups based on lactation stages and ages of animals.

Within the framework of quantum refereed steering games, quantum steerability can be certified without any assumption on the underlying state nor the measurements involved. Such a scheme is termed the measurement-device-independent (MDI) scenario. Here, we introduce a measure of steerability in an MDI scenario, i.e., the result merely depends on the observed statistics and the quantum inputs. We prove that such a measure satisfies the convex steering monotone. Moreover, it is robust against not only measurement biases but also losses. We also experimentally estimate the amount of the measure with an entangled photon source. As two by-products, our experimental results provide lower bounds on an entanglement measure of the underlying state and an incompatible measure of the involved measurement. Our research paves a way for exploring one-side device-independent quantum information processing within an MDI framework.

Magnetic impurities on the surface of Rashba spin-orbit-coupled, but otherwise conventional, superconductors provide a promising way to engineer topological superconductors with Majorana bound states as boundary modes. In this work we show that the spin-polarization in the interior of both one-dimensional impurity chains and two-dimensional islands in these systems can be used to determine the superconducting topological phase, as it changes sign exactly at the topological phase transition. Thus, spin polarization offers an alternative method to detect the topological phase in magnetic impurity chains and islands deposited on conventional superconductors, beyond the zero-energy Majorana bound states.

Background: After the 1968 United Nations International Conference on Human Rights, access to family planning services became a human right. Such a service is of central importance to women's empowerment and is empirically needed to provide adequate healthcare. For registered refugees and asylum seekers in Germany complementary family planning services, including all forms of contraception, are free of charge. Yet, the success of these services remains unclear. The aim of this study is to describe the current reproductive health status of female refugees and to provide an initial overview of their existing unmet family planning and contraception needs.

Methods: Over the course of 2 years, from December 2015 to December 2017, a set of 50 female-only discussion groups were conducted in community shelters for registered refugees in Berlin. A total of 410 women between the ages of 14 and 74 participated. A convenience sampling strategy was then applied and a total of 307 semi-structured questionnaires covering 41 items related to demographic data and women's health were distributed to volunteering female participants over the age of 17. The statistical analysis of the questionnaires was performed using SPSS (IBM, PASW, Version 24). P-values less than or equal to 0.05 were considered statistically significant.

Results: Of the 307 participants, the majority were from Syria and Afghanistan (30% respectively). The mean age was 33 years (range: 18-63). On average, each woman had 2.5 births (range: 0-10). Twenty-four women (8%) were pregnant and fifty-four of the women (18%) were trying to become pregnant. The majority of women were classified as "requiring contraception" (n = 195; 63%) of which 183 gave further information on if and how they used family planning methods. The calculated unmet need for family planning in this group was 47%. Of the remaining 53% of the women who used contraception, many utilised "traditional" methods (34% withdrawal method; 8% calendar method) which have a pearl index of 4-18 and can therefore be classified as rather insufficient birth control methods. Intrauterine contraceptive devices were used by 30%.

Conclusion: Our study revealed that despite the provision of complementary family planning services, there remains unmet family planning and education needs in the female refugee community in Berlin. This study indicates that there is a major access gap to these services. Further research needs to be carried out to evaluate the access gap and clearly identify and implement action plans to address possible causes such as language barriers, lack of childcare and traumatic experiences.

The introduction of biosimilars of biological agents for which the patents and exclusivity periods have expired is an attractive way of reducing healthcare spending through price competition with the reference product. In oncology, biosimilars of growth factors for supportive therapy were the pioneers; now, monoclonal antibody biosimilars are conquering the market. In Europe, this is currently limited to biosimilars of the monoclonal antibodies trastuzumab and rituximab. However, the pipeline is full and several monoclonal antibody biosimilars in oncology are now in late-stage development. We are expecting not only more biosimilar versions of the top 3 blockbuster monoclonal antibodies, trastuzumab, rituximab and bevacizumab, to enter the market; as patent expiration of multiple other cancer biologicals will occur in the next few years, the biosimilar landscape will become much more diversified. Several biosimilars of monoclonal antibodies used in targeted therapy such as cetuximab, pertuzumab, or denosumab are in early development.

Across group-living animals, linear dominance hierarchies lead to disparities in access to resources, health outcomes and reproductive performance. Studies of how dominance rank predicts these traits typically employ one of several dominance rank metrics without examining the assumptions each metric makes about its underlying competitive processes. Here, we compare the ability of two dominance rank metrics-simple ordinal rank and proportional or 'standardized' rank-to predict 20 traits in a wild baboon population in Amboseli, Kenya. We propose that simple ordinal rank best predicts traits when competition is density-dependent, whereas proportional rank best predicts traits when competition is density-independent. We found that for 75% of traits (15/20), one rank metric performed better than the other. Strikingly, all male traits were best predicted by simple ordinal rank, whereas female traits were evenly split between proportional and simple ordinal rank. Hence, male and female traits are shaped by different competitive processes: males are largely driven by density-dependent resource access (e.g. access to oestrous females), whereas females are shaped by both density-independent (e.g. distributed food resources) and density-dependent resource access. This method of comparing how different rank metrics predict traits can be used to distinguish between different competitive processes operating in animal societies.

We analyse the properties of an unbiased gradient estimator of the evidence lower bound (ELBO) for variational inference, based on the score function method with leave-one-out control variates. We show that this gradient estimator can be obtained using a new loss, defined as the variance of the log-ratio between the exact posterior and the variational approximation, which we call the log-variance loss. Under certain conditions, the gradient of the log-variance loss equals the gradient of the (negative) ELBO. We show theoretically that this gradient estimator, which we call VarGrad due to its connection to the log-variance loss, exhibits lower variance than the score function method in certain settings, and that the leave-one-out control variate coefficients are close to the optimal ones. We empirically demonstrate that VarGrad offers a favourable variance versus computation trade-off compared to other state-of-the-art estimators on a discrete variational autoencoder (VAE)

Background Effective and sustainable worm control in horses would benefit from detailed information about the current regional occurrence of tapeworms. Different diagnostic methods are currently available to detect Anoplocephala spp. infections in horses. However, the format as well as the sensitivity and specificity of the methods vary considerably.

Methods A coprological, serological and questionnaire study was conducted to investigate the prevalence and risk factors of tapeworm infections on 48 horse farms in the region of Berlin and Brandenburg, Germany. In total, faecal samples of 484 horses were analysed using the double centrifugation/combined sedimentation-flotation and mini-FLOTAC. Serum (n = 481) and saliva (n = 365) samples were analysed by ELISAs to determine antibody levels against Anoplocephala spp. 12/13 kDa excretory/secretory (E/S) antigens.

Results Cestode eggs were detected in 0.6% of faecal samples (farm prevalence 6.3%) without differences between the two methods. In contrast, antibodies against Anoplocephala spp. were detected in 16.2% (farm prevalence 52.1%) and in 29.5% (farm prevalence 75.7%) of the serum and saliva samples, respectively. Both ELISA based methods for detection of tapeworms reported a greater number of infected animals requiring treatment than were positively identified by coproscopy. Logistic regression analysis identified permanent pasture access, large pastures and regular pasture changes and high strongyle egg counts as risk factors for positive serum antibody responses to Anoplocephala spp. while last treatment with praziquantel was protective. Other protective factors were the presence of foals and high numbers of horses on the farm. Daily removal of faeces from the pasture and horse age did not have a significant effect.

Conclusions The findings of the present serological investigation indicate that tapeworm prevalence in Berlin/Brandenburg horse farms is much higher than would be anticipated by using conventional/coproscopic analyses. Moreover, the majority of tapeworm-positive horses had not received a cestocidal drug at their last treatment. Considering the already known low sensitivity of the coproscopic detection, the equine veterinary diagnostics can be enhanced by the use of antibody detection methods such as the saliva-based ELISA.