Dramatic changes in the structure of shallow-water marine benthic communities following the invasion by Rugulopteryx okamurae (Dictyotales, Ochrophyta) in Azores (NE Atlantic).

Biological invasions are considered one of the most important drivers of biodiversity loss. Here we use a before-after-control-impact (BACI) design to investigate the impact of Rugulopteryx okamurae on the structure of shallow-water marine benthic communities in São Miguel island, Azores. After its first appearance in 2019, R. okamurae has rapidly invaded much of the southern coast of the island, where it became the dominant algae. This was followed by significant changes in the structure of shallow-water marine benthic communities, with substantial losses of natural variability and species richness. Compared to before, there has been dramatic reductions in the abundances of articulated coralline algae, corticated algae and corticated foliose algae in invaded locations. These results highlight its highly invasive character, not seen with other, more well-known, invasive species. It remains to be investigated if its impacts persist throughout time and to quantify the functional consequences of such dramatic changes.

Evaluation of chickens infected with a recombinant virulent NDV clone expressing chicken IL4.

BACKGROUND: There is evidence that chicken IL4 (chIL4) functions similarly to its mammalian analogue by enhancing type 2 T helper (Th2) humoral immunity and promoting protection against parasitic infections; however, no studies have been performed to assess the effect of chIL4 on the pathogenesis of Newcastle disease (ND). To assess the role of chIL4 in velogenic NDV pathogenesis we created a vNDV infectious clone expressing chIL4. We hypothesized that co-expression of chIL4 during virus replication would result in decreased inflammation caused by the Th1 response and thereby increasing survival to challenge with vNDV. METHODS: To evaluate the effect of chIL4 during early infection with velogenic Newcastle disease virus (NDV) in chickens, recombinant NDV clones expressing either chIL4 (rZJ1-IL4) or a control expressing green fluorescent protein (rZJ1-GFP) were created by inserting an expression cassette in an intergenic region of the NDV genome. The pathogenesis of rZJ1-IL4 was assessed in 4-week-old specific pathogen free chickens. The extent of virus replication was evaluated by titration in mucosal secretions and immunohistochemistry in multiple tissues. Expression of chIL4 was confirmed in tissues using immunohistochemistry. RESULTS: Infection of birds with the rZJ1-IL4 resulted in successful viral replication in vivo and in vitro and generation of the chIL4 in tissues. All birds were clinically normal 2 DPI, with one bird in each group showing conjunctival swelling and enlarged spleens grossly. At 5 DPI, moderate or severe depression was observed in birds infected with rZJ1-GFP or rZJ1-IL4, respectively. Neurological signs and thymic atrophy were observed in one bird infected with rZJ1-IL4. Grossly, conjunctival swelling, mottled spleen and proventricular hemorrhages were observed at 5 DPI in one bird from each group. At 5 DPI, severe necrosis in the spleen, bursa and cecal tonsils were observed in birds infected with rZJ1-GFP, along with minimal evidence of chIL4 expression. In contrast, splenic atrophy, and moderate necrosis in the bursa and cecal tonsils were observed in birds infected with rZJ1-IL4. In addition, chIL4 signal was found in all tissues of rZJ1-IL4 birds at 5DPI. CONCLUSIONS: The production of chIL4 by a recombinant NDV strain resulted in the activation of the positive feedback loop associated with IL4 production. Insertion of chIL4 into NDV may decrease necrosis to lymphoid organs while increasing the severity of lymphoid atrophy and prolonged disease. However, with a low number of birds it is difficult to determine whether these results are significant to disease outcome.

Predicting the ecological impact of a recent range expansion in the structure of intertidal biofilms.

Predicting the consequences of recent changes in species distributional ranges is paramount. The trochid Phorcus sauciatus has recently colonised the Azores and is the only intertidal trochid in these islands. In this study we used experimental mesocosms to examine whether its addition to the remainder of the intertidal assemblage of grazers composed of littorinids and patellids affects the structure of epilithic biofilms growing on experimental plates. We also examined if its presence can compensate the loss of patellid limpets to simulate areas where these are chronically harvested. Results showed that when the native assemblage of grazers (littorinids and patellids) are present, the addition of P. sauciatus had little influence on the overall structure of epilithic biofilms. However, when patellids were absent, and in contrast to expectations, biofilm standing stock on experimental plates decreased significantly. Results suggest that patellids may negatively affect the foraging activities of P. sauciatus via interference competition and are further discussed in face of ecological knowledge of these organisms.

Marine algal flora of Santa Maria Island, Azores.

BACKGROUND: The algal flora of the Island of Santa Maria (eastern group of the Azores archipelago) has attracted interest of researchers on past occasions (Drouët 1866, Agardh 1870, Trelease 1897, Schmidt 1931, Ardré et al. 1974, Fralick and Hehre 1990, Neto et al. 1991, Morton and Britton 2000, Amen et al. 2005, Wallenstein and Neto 2006, Tittley et al. 2009, Wallenstein et al. 2009a, Wallenstein et al. 2010, Botelho et al. 2010, Torres et al. 2010, León-Cisneros et al. 2011, Martins et al. 2014, Micael et al. 2014, Rebelo et al. 2014, Ávila et al. 2015, Ávila et al. 2016, Machín-Sánchez et al. 2016, Uchman et al. 2016, Johnson et al. 2017, Parente et al. 2018). Nevertheless, the Island macroalgal flora is not well-known as published information reflects limited collections obtained in short-term visits by scientists. To overcome this, a thorough investigation, encompassing collections and presence data recording, was undertaken at both the littoral and sublittoral levels down to a depth of approximately 40 m, covering an area of approximately 64 km2. The resultant taxonomic records are listed in the present paper which also provides information on species ecology and occurrence around the Island, improving, thereby, the knowledge of the Azorean macroalgal flora at both local and regional scales. NEW INFORMATION: A total of 2329 specimens (including some taxa identified only to genus level) belonging to 261 taxa of macroalgae are registered, comprising 152 Rhodophyta, 43 Chlorophyta and 66 Ochrophyta (Phaeophyceae). Of these, 174 were identified to species level (102 Rhodophyta, 29 Chlorophyta and 43 Ochrophyta), encompassing 52 new records for the Island (30 Rhodophyta, 9 Chlorophyta and 13 Ochrophyta), 2 Macaronesian endemics (Laurencia viridis Gil-Rodríguez & Haroun; and Millerella tinerfensis (Seoane-Camba) S.M.Boo & J.M.Rico), 10 introduced (the Rhodophyta Acrothamnion preissii (Sonder) E.M.Wollaston, Antithamnion hubbsii E.Y.Dawson, Asparagopsis armata Harvey, Bonnemaisonia hamifera Hariot, Melanothamnus harveyi (Bailey) Díaz-Tapia & Maggs, Scinaia acuta M.J.Wynne and Symphyocladia marchantioides (Harvey) Falkenberg; the Chlorophyta Codium fragile subsp. fragile (Suringar) Hariot; and the Ochrophyta Hydroclathrus tilesii (Endlicher) Santiañez & M.J.Wynne, and Papenfussiella kuromo (Yendo) Inagaki) and 18 species of uncertain status (11 Rhodophyta, 3 Chlorophyta and 4 Ochrophyta).

Marine algal flora of São Miguel Island, Azores.

BACKGROUND: The macroalgal flora of the Island of São Miguel (eastern group of the Azores Archipelago) has attracted the interest of many researchers in the past, the first publications going back to the nineteenth century. Initial studies were mainly taxonomic, resulting in the publication of a checklist of the Azorean benthic marine algae. Later, the establishment of the University of the Azores on the Island permitted the logistic conditions to develop both temporal studies and long-term research and this resulted in a significant increase on research directed at the benthic marine algae and littoral communities of the Island and consequent publications.Prior to the present paper, the known macroalgal flora of São Miguel Island comprised around 260 species. Despite this richness, a significant amount of the research was never made public, notably Masters and PhD theses encompassing information regarding presence data recorded at littoral and sublittoral levels down to a depth of approximately 40 m around the Island and the many collections made, which resulted in vouchers deposited in the AZB Herbarium Ruy Telles Palhinha and the LSM- Molecular Systematics Laboratory at the Faculty of Sciences and Technology of the University of the Azores.The present publication lists the macroalgal taxonomic records, together with information on their ecology and occurrence around São Miguel Island, improving the knowledge of the Azorean macroalgal flora at local and regional scales. NEW INFORMATION: A total of 12,781 specimens (including some identified only to genus) belonging to 431 taxa of macroalgae are registered, comprising 284 Rhodophyta, 59 Chlorophyta and 88 Ochrophyta (Phaeophyceae). Of these, 323 were identified to species level (212 Rhodophyta, 48 Chlorophyta and 63 Ochrophyta), of which 61 are new records for the Island (42 Rhodophyta, 9 Chlorophyta and 10 Ochrophyta), one an Azorean endemic (Predaea feldmannii subsp. azorica Gabriel), five are Macaronesian endemisms (the red algae Botryocladia macaronesica Afonso-Carrillo, Sobrino, Tittley & Neto, Laurencia viridis Gil-Rodríguez & Haroun, Millerella tinerfensis (Seoane-Camba) S.M.Boo & J.M.Rico, Phyllophora gelidioides P.Crouan & H.Crouan ex Karsakoff and the green alga Codium elisabethiae O.C.Schmidt), 19 are introduced species (15 Rhodophyta, two Chlorophyta and two Ochrophyta) and 32 are of uncertain status (21 Rhodophyta, five Chlorophyta and six Ochrophyta).

Marine algal flora of Flores and Corvo Islands, Azores.

BACKGROUND: The algal flora of the western group of the Azores archipelago (Islands of Flores and Corvo) has attracted the interest of many researchers on numerous past occasions (such as Drouet 1866, Trelease 1897, Gain 1914, Schmidt 1929, Schmidt 1931, Azevedo et al. 1990, Fralick and Hehre 1990, Neto and Azevedo 1990, Neto and Baldwin 1990, Neto 1996, Neto 1997, Neto 1999, Tittley and Neto 1996, Tittley and Neto 2000, Tittley and Neto 2005, Tittley and Neto 2006, Azevedo 1998, Azevedo 1999, Tittley et al. 1998, Dionísio et al. 2008, Neto et al. 2008). Despite this interest, the macroalgal flora of the Islands cannot be described as well-known with the published information reflecting limited collections preformed in short-term visits by scientists. To overcome this, a thorough investigation, encompassing collections and presence data recording, has been undertaken for both the littoral and sublittoral regions, down to a depth of approximately 40 m, covering a relatively large area on both Islands (approximately 143 km2 for Flores and 17 km2 for Corvo).This paper lists the resultant taxonomic records and provides information on species ecology and occurrence around both these Islands, thereby improving the knowledge of the Azorean macroalgal flora at both local and regional scales. NEW INFORMATION: For the Island of Flores, a total of 1687 specimens (including some taxa identified only to genus level) belonging to 196 taxa of macroalgae are registered, comprising 120 Rhodophyta, 35 Chlorophyta and 41 Ochrophyta (Phaeophyceae). Of these taxa, 128 were identified to species level (80 Rhodophyta, 22 Chlorophyta and 26 Ochrophyta), encompassing 37 new records for the Island (20 Rhodophyta, 6 Chlorophyta and 11 Ochrophyta); two Macaronesian endemics (Laurencia viridis Gil-Rodríguez & Haroun and Millerella tinerfensis (Seoane-Camba) S.M.Boo & J.M.Rico); six introduced (the Rhodophyta Asparagopsis armata Harvey, Neoizziella divaricata (C.K.Tseng) S.-M.Lin, S.-Y.Yang & Huisman and Symphyocladia marchantioides (Harvey) Falkenberg; the Chlorophyta Codium fragile subsp. fragile (Suringar) Hariot; and the Ochrophyta Hydroclathrus tilesii (Endlicher) Santiañez & M.J.Wynne and Papenfussiella kuromo (Yendo) Inagaki); and 14 species of uncertain status (10 Rhodophyta, two Chlorophyta and two Ochrophyta).For the Island of Corvo, a total of 390 specimens distributed in 56 taxa of macroalgae are registered, comprising 30 Rhodophyta, nine Chlorophyta and 17 Ochrophyta (Phaeophyceae). Whilst a number of taxa were identified only to the genus level, 43 were identified to species level (22 Rhodophyta, eight Chlorophyta and 13 Ochrophyta), comprising 22 new records for the Island (nine Rhodophyta, four Chlorophyta and nine Ochrophyta), two introduced species (the Rhodophyta Asparagopsis armata and the Chlorophyta Codium fragile subsp. fragile and seven species of uncertain status (five Rhodophyta and two Ochrophyta).

Marine algal flora of Graciosa Island, Azores.

BACKGROUND: The macroalgal flora of Graciosa (central group of Azores archipelago) is poorly known, with only 116 species recorded so far (authors personal data). The published information reflects occasional collections from sporadic field visits to the Island. To overcome this, a thorough investigation under the Expedition "GRACIOSA/2004", the Campaigns "PADEL/2006", "MACROBIOLMOL/2014" and "PIMA-BALA/2017" involving sample collecting and presence data recording, was undertaken over an area of 19 km2 encompassing littoral and sublittoral levels down to about 40 m around the Island. This paper lists the taxonomic records and provides information on species ecology and occurrence around the Island improving the knowledge of the Azorean macroalgal flora at both local and regional scales. NEW INFORMATION: A total of 1692 specimens belonging to 250 taxa of macroalgae (and including 55 taxa identified only at the genus level) are registered, comprising 166 Rhodophyta, 36 Chlorophyta and 48 Ochrophyta (Phaeophyceae). From these, 195 are identified to the species level (126 Rhodophyta, 31 Chlorophyta and 38 Ochrophyta) and comprise 156 native, 20 of uncertain origin and 14 introductions to the Island. Predaea feldmannii subsp. azorica Gabriel is an Azorean endemic, whereas Codium elisabethiae O.C. Schmidt, Botryocladia macaronesica Afonso-Carrillo, Sobrino, Tittley & Neto, Phyllophora gelidioides P.Crouan & H.Crouan ex Karsakoff and Laurencia viridis Gil-Rodríguez & Haroun represent Macaronesian endemics. Seventy-nine species are newly recorded to the algal flora of the Island.

Marine algal (seaweed) flora of Terceira Island, Azores.

BACKGROUND: As for many other Azorean Islands, the macroalgal flora of Terceira (belonging to the central group of the archipelago) is poorly known, the published information reflecting occasional collections of sporadic visitors to the island. In order to overcome this and contribute to improve the knowledge of Azorean macroalgal flora at both local and regional scales, a thorough investigation was conducted. Both collections and presence data recordings were undertaken at the littoral and sublittoral levels down to approximately 40 m around the island, covering a total area of approximately 49 km2. This paper lists the taxonomic records and provides information on each species' ecology and occurrence on the Island's littoral. NEW INFORMATION: A total of 418 specimens (including taxa identified only to genus level) belonging to 147 taxa of macroalgae, comprising 95 Rhodophyta, 33 Chlorophyta and 19 Ochrophyta (Phaeophyceae) are registered. Of these, 113 were identified to species level (73 Rhodophyta, 24 Chlorophyta and 16 Ochrophyta), encompassing 35 new records for the Island (27 Rhodophyta, 6 Chlorophyta and 2 Ochrophyta). Most species are native, including the Macaronesian endemisms Codium elisabethiae O.C.Schmidt, Millerella tinerfensis (Seoane-Camba) S.M.Boo & J.M.Rico and Phyllophora gelidioides P.Crouan & H.Crouan ex Karsakoff. Eight species are introduced and 15 have uncertain origin.

Marine algal flora of Formigas Islets, Azores.

BACKGROUND: The oldest reference to marine life in Formigas Islets (oriental group of the Azores archipelago) goes back to the 16th century. Nevertheless, their macroalgal flora is poorly known, the published information mainly resulting from occasional collections of sporadic visitors. To overcome this and contribute to the knowledge of Azorean macroalgal flora at both local and regional scales, a thorough investigation was conducted in 1990 and 1991 under two expeditions promoted by the Marine Biology Research Group of the Department of Biology, University of the Azores. Collections and presence data recordings were undertaken at the littoral and sublittoral levels down to approximately 40 m, in an area of approximately 0.04 km2. This paper lists the taxonomic records and provides information regarding each species' ecology and occurrence on the Islets' littoral. NEW INFORMATION: A total of 320 specimens are registered (including taxa identified only at generic level) belonging to 90 taxa of macroalgae, from which 70 were diagnosed at species level. The confirmed species comprise 39 Rhodophyta, 12 Chlorophyta and 19 Ochrophyta (Phaeophyceae), distributed in 22 orders (13 Rhodophyta, 3 Chlorophyta and 6 Ochrophyta) and 37 families (24 Rhodophyta, 6 Chlorophyta and 7 Ochrophyta). Sixty-one species represent new records for the Islets, from which Botryocladia macaronesica Afonso-Carrillo, Sobrino, Tittley & Neto and Laurencia viridis Gil-Rodriguez & Haroun are Macaronesian endemisms. Most species are native to the Azores, but six have an uncertain origin and four are introduced (the Rhodophyta Asparagopsis armata Harvey; Laurencia dendroidea J.Agardh; Neoizziella divaricata (C.K.Tseng) S.-M.Lin, S.-Y.Yang & Huisman and the Ochrophyta Hydroclathrus tilesii (Endlicher) Santiañez & M.J.Wynne).

Marine algal flora of Pico Island, Azores.

BACKGROUND: The seaweed flora of Pico Island (central group of the Azores archipelago) has attracted interest of researchers on past occasions. Despite this, the macroalgal flora of the island cannot be considered well-known as published information reflects only occasional collections. To overcome this, a thorough investigation encompassing collections and presence data recording was undertaken. Research under the Campaigns "AÇORES/89", "PICO/91", "PICOBEL/2007" and "LAUMACAT/2011" covered a relatively large area (approximately 39 km2) around the island, encompassing the littoral and sublittoral levels down to about 40 m around the Island.This paper improves the knowledge of the Azorean macroalgal flora at local and regional scales by listing taxonomic records and providing information on the ecology and occurrence of each species present on the Island's littoral. NEW INFORMATION: A total of 4043 specimens (including taxa identified only to genus level) belonging to 303 taxa of macroalgae are registered, comprising 197 Rhodophyta, 53 Chlorophyta and 53 Ochrophyta (Phaeophyceae). From these, 225 were identified to species level (142 Rhodophyta, 41 Chlorophyta and 42 Ochrophyta), encompassing 110 new records for the island (69 Rhodophyta, 20 Chlorophyta and 21 Ochrophyta), three Macaronesian endemisms (Botryocladia macaronesica Afonso-Carillo, Sobrino, Tittley & Neto; Laurencia viridis Gil-Rodríguez & Haroun; Codium elisabethiae O. C. Schmidt), 14 introduced and 25 species with an uncertain status.

The Azorean edible abalone Haliotis tuberculata, an alternative heavy metal-free marine resource?

Abalones are considered a delicacy and exploited for food worldwide. In many places, overfishing has led to the decimation of wild stocks and many are now reared in aquaculture systems. In the Azores, there is no tradition of eating abalones and Haliotis tuberculata stocks are still virtually untouched. However, as tourism in the islands grows and the stocks of other shellfish diminish, there is an increasing pressure to find alternative edible resources, leading to a rising interest in abalones. According to previous studies performed in the region, other edible species, including the local highly appreciated limpets and the giant barnacle, present high concentration levels of some heavy metals, which has been attributed to the volcanic origin of the islands. Here we analysed the metal content in the edible tissue of Haliotis tuberculata from São Miguel Island, Azores. The potential human health risks due to its consumption was assessed by estimating the average daily intake (EDI) and target hazard quotient (THQ) of metals. Similarly to other organisms in the Azores, abalones have higher than normal levels of some heavy metals, particularly cadmium, reflecting a local natural source that should be closely monitored from a public health point of view.

Presence of Newcastle disease viruses of sub-genotypes Vc and VIn in backyard chickens and in apparently healthy wild birds from Mexico in 2017.

Virulent Newcastle disease viruses (NDV) have been present in Mexico since 1946, and recently, multiple outbreaks have been reported in the country. Here, we characterized eleven NDV isolated from apparently healthy wild birds and backyard chickens in three different locations of Jalisco, Mexico in 2017. Total RNA from NDV was reverse-transcribed, and 1285 nucleotides, which includes 3/4 of the fusion gene, was amplified and sequenced using a long-read MinION sequencing method. The sequences were 99.99-100% identical to the corresponding region obtained using the Illumina MiSeq. Phylogenetic analysis using MinION sequences demonstrated that nine virulent NDV from wild birds belonged to sub-genotypes Vc and VIn, and two backyard chicken isolates were of sub-genotype Vc. The sub-genotype Vc viruses had nucleotide sequence identity that ranged from 97.7 to 98% to a virus of the same sub-genotype isolated from a chicken in Mexico in 2010. Three viruses from pigeons had 96.3-98.7% nucleotide identity to sub-genotype VIn pigeon viruses, commonly referred to as pigeon paramyxovirus, isolated in the USA during 2000-2016. This study demonstrates that viruses of sub-genotype Vc are still present in Mexico, and the detection of this sub-genotype in both chickens and wild birds suggests that transmission among these species may represent a biosecurity risk. This is the first detection and complete genome sequencing of genotype VI NDV from Mexico. In addition, the utilization of an optimized long-read sequencing method for rapid virulence and genotype identification using the Oxford nanopore MinION system is demonstrated.

Epidemiology, control, and prevention of Newcastle disease in endemic regions: Latin America.

Newcastle disease (ND) infects wild birds and poultry species worldwide, severely impacting the economics of the poultry industry. ND is especially problematic in Latin America (Mexico, Colombia, Venezuela, and Peru) where it is either endemic or re-emerging. The disease is caused by infections with one of the different strains of virulent avian Newcastle disease virus (NDV), recently renamed Avian avulavirus 1. Here, we describe the molecular epidemiology of Latin American NDVs, current control and prevention methods, including vaccines and vaccination protocols, as well as future strategies for control of ND. Because the productive, cultural, economic, social, and ecological conditions that facilitate poultry endemicity in South America are similar to those in the developing world, most of the problems and control strategies described here are applicable to other continents.

Pathology and Distribution of Velogenic Viscerotropic Newcastle Disease Virus in the Reproductive System of Vaccinated and Unvaccinated Laying Hens (Gallus gallus domesticus) by Immunohistochemical Labelling.

This study investigated the pathological changes in the reproductive system of laying hens that lead to the poor egg production and quality in Newcastle disease (ND) and the distribution of the virus in the system. Two hundred and forty Isa-Brown pullets were divided randomly into vaccinated and unvaccinated groups (n = 120 each). The vaccinated group was given Hitchner B1 vaccine at 1 day of age, La Sota vaccine at 4 weeks of age and Komarov vaccine at 9 and 16 weeks of age. At the peak of egg production, the laying hens (32 weeks old) were assigned randomly into four groups (n = 60): VC, vaccinated with ND vaccines and inoculated intramuscularly with velogenic viscerotropic ND virus (vvNDV); VU, vaccinated unchallenged; UC, unvaccinated challenged; and UU, unvaccinated unchallenged. UC hens showed depression, diarrhoea and later torticollis. Mortality in UC hens was 90%. VC hens showed mild anorexia. The body weights of the UC hens were significantly (P <0.05) lower than those of UU hens. VC and UC hens showed a significant (P <0.05) drop in egg production. Only UC hens produced abnormal eggs and initially had swollen, oedematous, hyperaemic oviducts followed by atrophy and shortening of the reproductive tract with atresia of the ovarian follicles. The histopathological changes were of necrosis of the epithelium and secretory glands. VC hens showed mild inflammatory changes in the oviduct. Immunohistochemical labelling showed extensive presence of the virus in the ovary, infundibulum, magnum, isthmus, uterus and vagina of UC hens and in the ovary of VC hens. These changes will be the cause of serious egg production problems, especially in vaccinated layers in countries where vvNDV is enzootic.

Natural Infections With Pigeon Paramyxovirus Serotype 1: Pathologic Changes in Eurasian Collared-Doves ( Streptopelia decaocto) and Rock Pigeons ( Columba livia) in the United States.

Pigeon paramyxovirus serotype 1 (PPMV-1) is a globally distributed, virulent member of the avian paramyxovirus serotype 1 serogroup that causes mortality in columbiformes and poultry. Following introduction into the United States in the mid-1980s, PPMV-1 rapidly spread causing numerous mortality events in Eurasian collared-doves ( Streptopelia decaocto) (ECDOs) and rock pigeons ( Columba livia) (ROPIs). The investigators reviewed pathological findings of 70 naturally infected, free-ranging columbiforms from 25 different mortality events in the United States. Immunohistochemistry targeting PPMV-1 nucleoprotein was used to determine the tissue distribution of the virus in a subset of 17 birds from 10 of the studied outbreaks. ECDOs (61 birds) and ROPIs (9 birds) were the only species in which PPMV-1-associated disease was confirmed by viral isolation and presence of histologic lesions. Acute to subacute tubulointerstitial nephritis and necrotizing pancreatitis were the most frequent histologic lesions, with immunolabeling of viral antigen in renal tubular epithelial cells and pancreatic acinar epithelium. Lymphoid depletion of bursa of Fabricius and spleen was common, but the presence of viral antigen in these organs was inconsistent among infected birds. Hepatocellular necrosis was occasionally present with immunolabeling of hypertrophic Kupffer cells, and immunopositive eosinophilic intracytoplasmic inclusion bodies were present in hepatocytes of 1 ECDO. Immunopositive lymphocytic choroiditis was present in 1 ECDO, while lymphocytic meningoencephalitis was frequent in ROPIs in absence of immunolabeling. This study demonstrates widespread presence of PPMV-1 antigen in association with histologic lesions, confirming the lethal potential of this virus in these particular bird species.

Pathogenesis of New Strains of Newcastle Disease Virus From Israel and Pakistan.

In the past few years, Newcastle disease virus (NDV) strains with epizootic characteristics belonging to subgenotypes VIIi and XIIIb emerged in the Middle East and Asia. In this study, 2 NDV strains-1 representative of subgenotype VIIi isolated in Israel (Kvuzat/13) and 1 representative of subgenotype XIIIb isolated in Pakistan (Karachi/07)-were characterized by intracerebral pathogenicity index and detailed clinicopathologic assessment. The intracerebral pathogenicity index values for Kvuzat/13 and Karachi/07 were 1.89 and 1.85, respectively, classifying these strains as virulent by international standards. In 4-week-old White Leghorn chickens, both strains caused 100% mortality within 4 (Kvuzat/13) and 5 (Karachi/07) days postinfection. Histopathology and immunohistochemistry for NDV nucleoprotein showed that both strains had wide systemic distribution, especially targeting lymphoid organs and mucosa-associated lymphoid tissues in the respiratory and intestinal tracts. Results of the animal experiment confirm that both Kvuzat/13 and Karachi/07 are highly virulent and behaved as velogenic viscerotropic NDV strains.

Neuropathogenic Capacity of Lentogenic, Mesogenic, and Velogenic Newcastle Disease Virus Strains in Day-Old Chickens.

Strains of Newcastle disease virus (NDV) have different abilities to elicit neurologic signs. To determine the capacity of different NDV strains to replicate and cause lesions in the brain, independently of their peripheral replication, 1-day-old chickens were inoculated in the subdural space with 7 NDV strains of different virulence (4 velogenic, 2 mesogenic, 1 lentogenic). Velogenic strains induced severe necrotizing and heterophilic ventriculitis and meningitis, as well as edema of the neuroparenchyma, and replicated extensively in the nervous tissue by day 2 postinfection, as demonstrated by immunohistochemistry, when all infected birds died. Clinical signs, microscopic lesions, and viral replication were delayed (days 3 and 4 postinfection) with mesogenic strains. Velogenic and mesogenic NDV strains replicated mainly in neurons, and immunolabeling was first detected in surface-oriented areas (periventricular and submeningeal), possibly as a reflection of the inoculation route. The lentogenic NDV strain did not cause death of infected birds; replication was confined to the epithelium of the ependyma and choroid plexuses; and lesions consisted of lymphoid aggregates limited to the choroid plexuses. Results show that extensive NDV replication in the brain is typical of velogenic and mesogenic, but not lentogenic, NDV strains. In addition, this study suggests that differences in the rate of NDV replication in nervous tissue, not differences in neurotropism, differentiate velogenic from mesogenic NDV strains. This study indicates that intracerebral inoculation might be used as an effective method to study the mechanisms of NDV neuropathogenesis.

Pathologic characterization of genotypes XIV and XVII Newcastle disease viruses and efficacy of classical vaccination on specific pathogen-free birds.

To characterize the clinicopathologic features of recently described genotypes of Newcastle disease virus (NDV), 1 representative strain of genotype XIV and 2 of genotype XVII, all isolated from West Africa, were used to infect groups of ten 4-week-old specific pathogen-free chickens. The pathobiology of these 3 strains was compared to a South African NDV strain classified within genotype VII. All chickens infected with the 4 viruses died or were euthanized by day 4 postinfection due to the severity of clinical signs. Gross and histologic lesions in all infected chickens included extensive necrosis of lymphoid tissues (thymus, spleen, bursa of Fabricius, cecal tonsils, gut-associated lymphoid tissue), gastrointestinal necrosis and hemorrhages, and severe hemorrhagic conjunctivitis. Immunohistochemical staining revealed systemic viral distribution, and the most intense staining was in the lymphoid organs. Results demonstrate that the 3 West African strains from the previously uncharacterized genotypes XIV and XVII are typical velogenic viscerotropic NDV strains with lesions similar to the South African strain. Under experimental conditions, QV4 and LaSota NDV vaccine strains successfully protected chickens from morbidity and mortality against the genotype VII and one genotype XVII NDV strain, with no significant differences in the amount of virus shed when 2 vaccine schemes were compared.

Co-infection of mallards with low-virulence Newcastle disease virus and low-pathogenic avian influenza virus.

Waterfowl are considered the natural reservoir of low-virulence Newcastle disease viruses (loNDVs) and low-pathogenic avian influenza viruses (LPAIVs). The objective of this study was to investigate the effect of co-infections with loNDV and LPAIV on the infectivity and excretion of these viruses in mallards. One-month-old mallards were inoculated intranasally with 10(6) median embryo infectious doses of a wild-bird-origin loNDV and A/Mallard/MN/199106/99 (H3N8) LPAIV on the same day or received the LPAIV 2 or 5 days after loNDV inoculation. All mallards became infected with both viruses based on detection of seroconversion and viral shedding. Co-infection resulted in a higher number of cloacal swabs detected positive for LPAIV and a lower number of cloacal swabs detected positive for loNDV in some groups, although differences between groups were not statistically significant. Co-infection did not affect replication of LPAIV in epithelial cells of the lower intestine and bursa of Fabricius. In summary, the results of this study indicate that co-infection with LPAIV and loNDV does not affect the ability of mallards to be infected with either virus although it may have minimal effects on patterns (source and timing) of viral shedding.

Macular thickness measurements with frequency domain-OCT for quantification of axonal loss in chronic papilledema from pseudotumor cerebri syndrome.

PURPOSE: To evaluate the ability of frequency domain-optical coherence tomography (FD-OCT)-measured macular thickness parameters to differentiate between eyes with resolved chronic papilledema and healthy eyes and to evaluate the correlation between FD-OCT measures and visual field (VF) loss on standard automated perimetry (SAP). METHODS: Fifty-two eyes from 29 patients suffering from pseudotumor cerebri syndrome (PTC) and 62 eyes from 31 normal controls underwent FD-OCT scanning and ophthalmic evaluation including VF with SAP. All patients had previously been submitted to treatment of PTC and had clinically resolved papilledema and stable VF for at least 6 months before the study. Macular and peripapillary retinal nerve fiber layer (RNFL) thickness measurements were determined for both groups. Comparisons were made using Generalized Estimated Equations. Correlations between FD-OCT and VF measurements were verified. RESULTS: In eyes with resolved papilledema, the macular thickness parameters corresponding to the inner and outer superior, temporal, inferior and nasal segments, average macular thickness and most RNFL thickness measurements were significantly reduced when compared with controls. The discrimination ability was similar for macular thickness measurements and RNFL thickness measurements. Both sets of OCT measurements correlated well with VF sensitivity loss. CONCLUSIONS: Eyes with resolved chronic papilledema show a significant reduction in macular thickness, which is well correlated with the severity of VF loss. Macular thickness measurements can potentially be used to estimate and monitor the amount of ganglion cell loss in eyes with papilledema from patients with PTC.