Blood biomarkers and levels of major pollutants in eggs and feathers were used to determine pollution effects in nestlings of the Purple Heron Ardea purpurea and the Little Egret Egretta garzetta, sampled on three Ebro River (NE Spain) areas: a reference site, a site affected by the effluents of a chlor-alkali industry and the river Delta. The two impacted heron populations showed mutually different pollutant and response patterns, suggesting different sources of contamination. In the population nesting near the chlor-alkali plant, elevated levels of hexachlorobenzene (HCB) and polychlorobiphenyls (PCBs) in eggs, and mercury in feathers in A. purpurea chicks were related with reduced blood antioxidant defenses and increased levels of micronuclei. In Ebro Delta, high levels of plasmatic lactate dehydrogenase in A. purpurea chicks and high frequency of micronuclei in blood of both species were tentatively associated with intensive agricultural activities taking place in the area. These results provide the first evidence of a biological response in heron chicks to the release of pollutants at a chlor-alkali plant. High levels of organochlorine and mercury levels in eggs and feathers were related with altered blood biomarkers of heron nesting chicks.

Elasmobranchs can bioaccumulate considerable amounts of persistent organic pollutants (POPs) and utilize several reproductive strategies thereby influencing maternal transfer of contaminants. This study provides preliminary data on the POP transfer from pregnant females to offspring of three species (Atlantic stingrays, bonnethead, blacktip sharks) with different reproduction modes (aplacental, placental viviparity). Polychlorinated biphenyl (PCB) levels were generally higher than any other POPs. Stingrays and blacktip shark embryos contained the lowest POP concentrations while bonnetheads and the blacktip adult female had the highest concentrations. Results suggest that POPs are more readily transferred from the mother to the embryo compared to what is transferred to ova in stingrays. Statistically significant differences in levels of selected POPs were found between embryos from the left and right uterus within the same litter as well as between female and male embryos within the same litter for bonnetheads, but not for the blacktip sharks.

Although exploited populations of elasmobranchs may be able to recover from fishing pressure, there is little information regarding the Barndoor Skate's ability to cope with other anthropogenic stressors such as organic contaminants (OCs). Legacy OCs were measured in liver, muscle and ova from fourteen Barndoor Skates with mature skates having significantly greater mean concentrations of OCs than immature skates, demonstrating bioaccumulation with age. Using Toxic Equivalency Factors, skates were found to have levels of PCBs that have been shown to elicit negative physiological responses in other fishes and these results highlight the need for future studies to investigate the potential impacts that bioaccumulated organic contaminants have on the recovery and conservation of this vulnerable species.

Continuous monitoring and early warning together represent an important mitigation strategy for harmful algal blooms (HAB). The coast of Texas experiences periodic blooms of three HAB dinoflagellates: Karenia brevis, Dinophysis ovum, and Prorocentrum texanum. A plankton image data set acquired by an Imaging FlowCytobot over a decade of operation was used to train and evaluate two new automated image classifiers. A 112 class, random forest classifier (RF_112) and a 112 class, convolutional neural network classifier (CNN_112) were developed and compared with an existing, 54 class, random forest classifier (RF_54) already in use as an early warning notification system. Both 112 class classifiers exhibited improved performance over the RF_54 classifier when tested on three different HAB species with the CNN_112 classifier producing fewer false positives and false negatives in most of the cases tested. For K. brevis and P. texanum, the current threshold of 2 cells.mL⁻¹ was identified as the best threshold to minimize the number of false positives and false negatives. For D. ovum, a threshold of 1 cell.mL⁻¹ was found to produce the best results with regard to the number of false positives/negatives. A lower threshold will result in earlier notification of an increase in cell concentration and will provide state health managers with increased lead time to prepare for an impending HAB.

In this study, we have evaluated the efficacy of propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) to differentiate between viable and non-viable Ancylostoma caninum ova. The newly developed method was validated using raw wastewater seeded with known numbers of A. caninum ova. Results of this study confirmed that PMA-qPCR has resulted in average of 88 % reduction (P < 0.05) in gene copy numbers for 50 % viable +50 % non-viable when compared with 100 % viable ova. A reduction of 100 % in gene copies was observed for 100 % non-viable ova when compared with 100 % viable ova. Similar reductions (79–80 %) in gene copies were observed for A. caninum ova-seeded raw wastewater samples (n = 18) collected from wastewater treatment plants (WWTPs) A and B. The newly developed PMA-qPCR method was applied to determine the viable ova of different helminths (A. caninum, A. duodenale, Necator americanus and Ascaris lumbricoides) in raw wastewater, human fecal and soil samples. None of the unseeded wastewater samples were positive for the above-mentioned helminths. N. americanus and A. lumbricoides ova were found in unseeded human fecal and soil samples. For the unseeded human fecal samples (1 g), an average gene copy concentration obtained from qPCR and PMA-qPCR was found to be similar (6.8 × 10⁵ ± 6.4 × 10⁵ and 6.3 × 10⁵ ± 4.7 × 10⁵) indicating the presence of viable N. americanus ova. Among the 24 unseeded soil samples tested, only one was positive for A. lumbricoides. The mean gene copy concentration in the positively identified soil sample was 1.0 × 10⁵ ± 1.5 × 10⁴ (determined by qPCR) compared to 4.9 × 10⁴ ± 3.7 × 10³ (determined by PMA-qPCR). The newly developed PMA-qPCR methods were able to detect viable helminth ova from wastewater and soil samples and could be adapted for health risk assessment.

Treatment of domestic wastewater in a pilot-scale upflow anaerobic hybrid (AH) reactor (0.9 m³) in combination with downflow hanging sponge (DHS) system (1.3 m³) was investigated. The combined system was operated at a hydraulic retention time (HRT) of 6.0 h for AH and 3.2 h for DHS system. The total process achieved a substantial reduction of CODₜₒₜₐₗ resulting in an average effluent concentration of only 39 ± 12 mg/l. Moreover, 90 ± 7 % of ammonia was eliminated in the DHS system. Nitrate and nitrite data revealed that 49 ± 3.2 % of the ammonia removal occurred through nitrification process. The removal efficiency of total coliform (TC), fecal coliform (FC), and fecal streptococci (FS) was relatively low in the AH reactor. The major portion of TC, FC, and FS was removed in the DHS system resulting to an average count of 1.7 × 10⁵ ± 1.1 × 10²/100 ml for TC, 7.1 × 10⁴ ± 1.2 × 10²/100 ml for FC, and 7.5 × 10⁴ ± 1.3 × 10²/100 ml for FS in the final effluent. Likely, the combined system was very efficient for the removal of protozoological species such as sarcodins (Entamoeba cysts), flagellates (Giardia cysts), and ciliates (Balantidium cysts). This was not the case for coccidia (Cryptosporidium oocysts), where 36.4 and 27.3 % were detected in the effluent of AH and DHS system, respectively. Only 10 % of intestinal nematode and cestode ova were recorded in the effluent of AH reactor and were completely removed in the DHS system.